ISSN 1022-7954, Russian Journal of Genetics, 2009, Vol. 45, No. 3, pp. 276–286. © Pleiades Publishing, Inc., 2009.

Original Russian Text © B.F. Chadov, E.V. Chadova, E.A. Khotskina, N.B. Federova, 2009, published in Genetika, 2009, Vol. 45, No. 3, pp. 318–329.

MOLECULAR GENETICS

Conditional Lethal Mutations Shift the Genome

from Stability to Instability
B. F. Chadov, E. V. Chadova, E. A. Khotskina, and N. B. Federova
Institute of Cytology and Genetics, Russian Academy of Sciences, Novosibirsk, 630090 Russia;
e-mail: chadov@bionet.nsc.ru
Received July 27, 2007; in final form, April 8, 2008

Abstract—The phenomenology of genomic destabilization is described in Drosophila melanogaster mutants

containing radiation-induced conditional dominant lethals in the X chromosome and in autosome 2. Destabili-
zation manifests itself as (1) the loss or decrease of lethality of previously lethal mutations; (2) the loss of
expression of visible dominant mutations in an opposite homolog; (3) chromosomal instability resulting in the
loss of the X chromosome in germline and somatic cells; (4) the occurrence of novel mutations (secondary
mutagenesis); (5) the occurrence of single and mass modifications; (6) disturbances in individual development
(formation of morphoses). The key event for the shift of the genome from the stable state into the unstable one
is the occurrence of a conditional dominant lethal mutation.
DOI: 10.1134/S102279540903003X

INTRODUCTION In addition to conditional lethality, these mutations

The rule in choosing a character for genetic works,
introduced by Mendel, is its stable expression (high
expressivity) and stable inheritance (complete pene-
trance). This rule followed from the fundamental idea
that a factor (gene) is an elementary unit of inheritance,
which is not affected by variable inner and outer envi-
ronmental factors. It seemed then that the rule pre-
scribed by Mendel provided the choice of true (geneti-
cally determined) characters and excluded transient
(non-genetic) characters determined by the action of
environmental conditions.
The Mendelian approach to the choice of characters
did not alter with the beginning of the mutation era in
genetics. Artificially induced mutations also were to
display high expressivity and complete penetrance. Ide-
ally, a genetic mutation in a homozygote should be
expressed in any individual of a given species in an
unlimited number of generations.
A fundamentally new approach to choose mutations
was proposed in 2000: a mutation should be expressed
in individuals of one genotype and not expressed in
individuals of another genotype [1, 2]. It was supposed
that through selection of such mutations it is possible to
identify genes responsible for the formation of charac-
ters of intraspecific similarity [3, 4]. Characters of this
category were not studied by classical genetics [5].
Three techniques were developed to produce lethal
mutations in Drosophila melanogaster. These muta-
tions were defined as conditional dominant lethals. In
individuals of one genotype such mutations manifested
themselves as dominant lethals, and in individuals of
some other genotype they were not lethal [1, 6, 7].
showed other features. They stopped being lethal in the
presence of chromosome rearrangements [3, 7] and
demonstrated the parental effect in their expression
[7−9]. Offsprings of mutants often had complex devel-
opmental defects (morphoses) [8, 10, 11]. The capacity
to form morphoses and the conditional expression of
mutations made it possible to consider that the identi-
fied genes belong to the group of regulatory genes con-
trolling ontogenesis [4, 11–13].
One more remarkable feature was demonstrated in
the course of maintenance of D. melanogaster mutant
cultures: the mutants appeared to be genetically unsta-
ble. The first discovered form of instability was the loss
of lethal expression of mutations [9]. Then other forms
were established [13]. This publication is devoted to the
description of various forms of instability in mutants
containing conditional dominant lethals.
MATERIALS AND METHODS
Mutations in the X chromosome and in autosome 2
were obtained in 2000–2001 [1–4] (Figs. 1, 2). They
were maintained in culture for five years. From the for-
mal genetic point of view, the mutations are conditional
dominant lethals. The indicator of the lethal action of
mutations in the X chromosome is the lack of daughters
in crosses of a mutant male with females of the yellow
strain (Fig. 1). The indicator of the lethal action of
mutations in autosome 2 is the lack of mutant off-
springs of both sexes in similar crosses (Fig. 2).
X-chromosome mutations (22 mutations) were main-
tained by two ways: in culture with attached-X chromo-
somes and in culture with a Muller-5 inversion in the
X chromosome (Fig. 3).

276
 CONDITIONAL LETHAL MUTATIONS SHIFT THE GENOME 277

3000 R 3000 R
X X Y ×
X Y ×

 In(2LR)Cy/2 2/2

×
X  2/2 ×
2/In(2LR)Cy
Individual X X Y Individual
testing  2/2
×
2/In(2LR)Cy
testing X X ×
X Y
of males of males
 2/2 ×
2/In(2LR)Cy
*
X X ×
X* Y

Normal: 2/2 Ë 2/In(2LR)Cy

F1 Normal: daughters X X sons X Y F1
Mutation [*]: 2/2 * Ë 2/In(2LR)Cy
Mutation: daughters X X* sons X Y

Fig. 2. Discovery of conditional dominant lethals in auto-

Fig. 1. Discovery of conditional dominant lethals in X chro-
mosome of Drosophila melanogaster. Males exposed to
gamma rays were crossed with females containing attached
X chromosomes. The sons from the progeny were individu-
ally crossed with yellow females. The sons with the
acquired X chromosome containing a dominant lethal pro-
duced no daughters in their progeny [3].
some 2 of Drosophila melanogaster. Males exposed to
gamma rays were crossed with females containing a com-
plex inversion with the dominat marker Curly in one of
autosomes 2. The sons containing this autosome together
with the irradiated autosome 2 were individually crossed
with yellow females (shading). The sons with the acquired
autosome 2 containing a dominant lethal produced only
Curly offsprings [11].

(a) (b) y
In(1)M-5 In(1)M-5 v f
+
 ×
×

+  y v f

In(1)M-5
 In(1)M-5

In(1)M-5
F1
 F1
+ +


In(1)M-5


+

Fig. 3. Two ways of maintenance of ontogene mutations in X chromosome: (a) in a heterozygote in females containing an inverted
Muller-5 chromosome (In(1)M-5) and a mutant X chromosome (+, heavy line) in culture with attached X chromosomes (y v f/y v f).
In the first case the mutant X chromosome is acquired by In(1)M-5/+ daughters and + sons. In(1)M-5/In(1)M-5 daughters and
In(1)M-5 sons do not acquire the mutant X chromosome. In the second case the mutant X chromosome is acquired by the sons and
is not acquired by the daughters.

Lethal mutations in autosome 2 (eight mutations) mosome contained visible dominant mutations Curly,
were maintained in a heterozygote with an inverted Bristle, and Lobe distinguished by stable and clear
chromosome In(2LR)Cy, Cy Bl L4 (Fig. 4). This chro- expression [14].

RUSSIAN JOURNAL OF GENETICS Vol. 45 No. 3 2009

278 CHADOV et al.
lv1 2 4 2
 In(2L+2R)Cy, Cy dp pr Bl cn L sp
l (2)
× appear in the progeny, although their proportion did not
lv1 2 4 2

In(2L+2R)Cy, Cy dp pr Bl cn L sp
l (2)
Phenotype of offsprings
normally:  and
Cy Bl L4
Variants: Cy Bl –8
4
L –1
Cy –1
4 –9
Bl L
4 –0
Cy L lethality with both ways of maintenance. For most of
Bl –1
Fig. 4. Nonexpression of the phenotype of dominant muta-
tions in autosome 2. Lethal mutations l(2) in autosome 2
were maintained in culture containing inverted autosome 2
with mutations Curly (Cy), Bristle (Bl), and Lobe (L4). All
three dominant mutations were normally expressed in the
offsprings, but in 20 cases one or two mutations lost their
expression.
Each culture was maintained in four or five vials
with a standard Drosophila medium. Once or twice a
month in the flies were transferred to a fresh medium.
Upon the replacement, the progeny was examined
under a binocular microscope. New phenotypic vari-
ants, including morphoses, were documented with a
digital videocamera. In the case of appearance of an
individual with an unusual phenotype it was crossed
with sibs to derive a new strain. A repeated manifesta-
tion of this phenotype was searched for in the next gen-
eration in the original culture that gave rise to the new
phenotype.
Checking of lethality was made by a repeated test
cross of males from the mutant cultures with yellow
females [1, 2].
RESULTS
“Delethalization”
The loss of the lethal action of mutations was dis-
covered accidentally in 2001 in experiments on deter-
mination of the frequency of dominant lethals in the
progeny of mutant males. Males from cultures 1, 3, 5,
27, and 33 maintained with attached-X chromosomes
began to produce daughters in crosses with yellow
females (Table 1). In succeeding years, lethality was
checked in all cultures and in larger samples of off-
springs. The five above-mentioned mutations tested in
2002 and 2004 were indeed found to be nonlethal.
RUSSIAN JOURNAL OF GENETICS
Three more mutations (29, 38, 41) lost their lethallity in
2002 and one (35), in 2004. In some cultures lethality
did not disapper but decreased. Daughters began to
reach 50% (7, 9, 10, 11). Of 22 lethal mutations nine
mutations, completely lost lethality in the period from
2000 to 2004, and four mutations became semilethals.
The loss of lethality (mutations 1–41) also occurred
in the cultures maintained in a heterozygote with a
Muller-5 chromosome (Table 1). For the same period of
time, eight mutations (3, 5, 9, 10, 11, 32, 34, 38) com-
pletely lost their lethality, and in other eight mutations
(6, 7, 8, 30, 31, 33, 35, 36) it decreased. The difference
in the ways of mutation maintenance certainly had its
influence on the process of “delethalization”. Lethals
were best preserved in the Muller-5 culture. Delethal-
ization of mutations 3, 5, 38 occurred with both ways of
maintenance, while mutations 2, 6, 30, 31 preserved
the mutations (16 out of 22), preservation of lethality
and its loss were not coordinated in the cultures with a
Muller-5 chromosome and with attached X chromo-
somes.
In 2002, after some cultures displayed the loss and a
sharp decrease of lethality, ten cultures maintained with
attached X chromosomes were duplicated using new
cultures obtained from males taken from corresponding
Muller-5 cultures preserving lethality of mutations.
By 2004, seven of ten mutations, now maintained with
attached X chromosomes, again lost the lethal action
(Table 2). Three mutations (27, 29, 41) remained lethal.
These mutations also remained lethal in the Muller-5
culture (Table 1). The data obtained indicate that in the
cultures the properties of mutations change depending
on a mutation itself and on its genomic surroundings.
Loss of Expression of a Dominant Mutation
in an Opposite Chromosome
Lethal mutations in autosome 2 maintained in a het-
erozygote with an inverted chromosome In(2LR)Cy,
Cy Bl L4 are characterized by the loss of expression of
dominant mutations Cy, Bl, and L4 in the inverted chro-
mosome (Fig. 4). The loss occurred in crosses aimed at
maintaining mutations and in crosses between mutant
cultures. During the first six months of mutation main-
tenance 20 cases of the loss of expression were
recorded (Fig. 4). In 17 cases, one marker was lost, and
in three cases two markers (individuals Cy, Bl, L4). The
reciprocal classes, markedly differ in size. This argues
against losses resulting from crossingover. Chromo-
somes with an altered set of dominant markers contin-
ued behaving as balancers of lethals in autosome 2,
indicating that the crossingover block caused by the
presence of the In(2LR)Cy inversion remained as
before. The process leading to the alteration of expres-
sion of a dominant mutation was autonomous for each
mutation. For this reason, the loss of one mutation was
most frequent, and the loss of two mutations occurred
Vol. 45 No. 3 2009
 CONDITIONAL LETHAL MUTATIONS SHIFT THE GENOME 279

Table 1. The loss of lethal expression of mutations obtained in 2000 (maintenanace in attached-X and Muller-5 cultures)
2000, attached-X 2001, attached-X 2002, attached-X 2004, attached-X 2004, Muller-5
Culture
no. total proportion total proportion total proportion total proportion total proportion
progeny of daughters progeny of daughters progeny of daughters progeny of daughters progeny of daughters
1 191 0.00 13 *0.46 199 *0.42 77 *0.52 77 0.06
2 435 0.00 4 0.00 259 0.02 36 0.03 89 0.08
3 180 0.00 20 *0.45 311 *0.43 95 *0.50 77 *0.48
5 303 0.02 33 *0.45 265 *0.60 83 *0.41 64 *0.50
6 283 0.02 2 0.00 111 0.02 39 0.05 31 **0.10
7 100 0.00 3 0.00 44 **0.27 63 *0.40 11 **0.18
8 216 0.07 5 0.00 90 0.09 49 **0.14 68 **0.19
9 529 0.00 7 0.00 169 **0.21 81 0.04 118 *0.54
10 297 0.04 7 0.00 69 **0.30 57 **0.26 79 *0.42
11 409 0.06 4 0.00 82 **0.18 55 **0.16 86 *0.46
26 89 0.01 – 0.00 175 0.07 40 0.02 – –
27 161 0.00 29 *0.69 113 *0.56 92 *0.49 55 0.04
29 76 0.00 4 0.00 171 *0.54 80 *0.51 34 0.06
30 115 0.00 8 0.00 109 0.02 71 0.00 60 **0.10
31 189 0.00 8 0.00 138 0.01 70 0.03 50 **0.10
32 198 0.00 4 0.00 74 0.00 53 0.02 43 *0.54
33 234 0.00 23 *0.52 214 *0.56 88 *0.51 48 **0.10
34 198 0.00 – 0.00 62 0.00 54 0.02 67 *0.45
35 115 0.04 12 0.00 162 **0.13 83 *0.48 44 **0.14
36 110 0.01 5 0.00 106 0.02 54 0.07 30 **0.27
38 84 0.01 3 0.00 80 *0.56 51 **0.33 84 *0.50
41 100 0.01 5 0.00 331 *0.49 106 *0.52 93 0.06
Notes: * Loss of lethality.
** Decrease of lethality.

less frequently. Cases when three mutations at once
were lost are not known. Individually, each of the muta-
tions happened to be lost, but with different frequen-
cies. In 11 cases, the Curly mutation was lost, the Lobe
mutation was lost in ten cases, and Bristle was lost in
two cases.
The frequency of the loss of expression of dominant
mutations was high. Six cases of nonexpression among
90 offsprings (6.7%) were found in crossing two lethal
cultures (37 and 53). No losses of the marker were
observed among 833 offsprings in a cross of yellow
females with mutant males (four cultures) regarded as
the control one. It may be thought that the loss occurs
when a mutation is in the mother. It can be stated with
assurance that the loss of expression of dominant muta-
tions in the inverted chromosome In(2LR)Cy, Cy Bl L4
is caused by the mutations under study, since not a sin-
gle case of the loss of expression of dominant markers
was noted within many decades that this chromosome
was maintained in laboratory cultures. In(2LR)Cy chro-
mosomes with the lost markers were introduced in cul-
ture and showed stable inheritance of this change. Two
variants of the loss of Curly, one of Lobe, and one of
Bristle are still being maintained in culture.
Chromosomal Instability
Females with a mutaion in one X chromosome and
with a Muller-5 inversion in the other gave an unex-
pected progeny in crosses with yellow males: patrocli-
nous yellow males (Table 3). Patroclinous males were
found in the progeny of 20 of 21 mutations; in 11 muta-
tions the proportion of patroclinous males was very
high, over 10%. The appearance of patroclinous males
points to the loss or nondisjunction of X chromosomes
in meiosis in a female [15]. The study of other lethal
mutations showed that both processes take place during
oogenesis of mutant females.
The loss of chromosomes also occurs in mitotically
dividing cells of mutants. Male tissue regions were
found in daughters of mutant females, for example, yel-
low regions against the grey background in y/+ females,

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280 CHADOV et al.

Table 2. The loss of lethal expression of mutations initially legged, (2) interrupted vein (18), (3) interrupted vein
maintained in Muller-5 culture and then in C(1)DX, y w f culture (30), and (4) bubbles on wings. In contrast to the
Male remaining three mutations, the phenotype short-legged
Daughters Sons Total Proportion was complex. It is composed of four components:
culture
+ yellow progeny of daughters (1) the absence of four paw segments on each leg;
no.
(2) obliquely cut wings; (3) the absence of the second
1 53 65 118 0.45 longitudinal vein on the wings; and (4) the presence of
3 44 83 127 0.35 a bubble on one or two wings. The mutations were
5 62 49 111 0.56 recessive, they were inherited as X-chromosome muta-
7 72 60 132 0.54 tions, and expressed in females with 100% penetrance.
11 33 34 67 0.49 Males carrying each of these mutations had the normal
phenotype. These mutations were named dimorphic
27* 0 58 58 0
because each of them had two phenotypically different
29* 3 66 69 0.04 forms of manifestation [11].
33 47 61 108 0.44
Besides, dimorphic mutations are not stable. Within
38 66 60 126 0.52
two years after they were obtained, derivatives of two
41* 4 47 51 0.08 types appeared in each of the cultures. Mutations of the
* Mutations with preserved lethality. first type are expressed in both sexes with 100% pene-
trance. Mutations of the second type are expressed in
both sexes, but with incomplete penetrance. The deriv-
eyes of the Bar and wa phenotypes in Bar/+ and wa /+ atives of the short-legged mutation preserve the previ-
females (Fig. 5). These cases are explained by the loss ous four-component phenotype in a female, and in a
of a normal X chromosome in somatic cells. The loss of male components 3 and 4 are expressed. In the short-
X chromosome in early embryogensis led to the forma- legged, a change of the mutation localization took place
tion of gynandromorphs, i. e., mosaics combining a part in the case of formation of a culture with the manifes-
of the body of female constitution and a part of the body
of male constitution (Fig. 5). tation of the mutation in individuals of both sexes. The
mutation turned from X-chromosomal into autosomal
(autosome 3). The localization change process was
Formation of Visible Mutations accompanied by the formation of new mutations that
Mutations with complete penetrance. Flies with were phenotypically similar to the known mutations
lethal mutations had the normal phenotype. In the dumpy and black. The genetic analysis shows the reces-
course of culture maintenance individuals with abnor- sive character of each of them and their localization in
mal phenotypes began to appear. Most of them corre- autosome 2. In the cultures of dimorphous mutations
sponded to phenotypes of known mutations and there periodically occurred new hereditable changes of
received their names in accordance to that. These were the phenotype.
(1) plexus, (2) dumpy; 3) brown; 4) radius incompletus;
5) forked; 6) balloon; 7) Bar, and flies with the wings Formation of a set of mutations in one genera-
directed at an angle to each other (“house” phenotype). tion or in a succession of generations. In one gen-
All mutations, except Bar, were recessive and had eration there appeared three individuals, each con-
100% penetrance in a homozygote. Their relation to the taining a complex of four mutations. Three are
known mutations with similar phenotypes remains to allelic to the known mutations in autosome 3 of
be studied. Drosophila: thread, scarled, and radius incomple-
Mutations with incomplete penetrance. They dif- tus. The fourth one is similar to the eyeless muta-
fered from the above-listed mutations in that part of tion in autosome 4. The complex of four mutations
mutation carriers had the normal phenotype. The pres- was found in several test tubes at once. Consecutive
ence of a mutation in them was revealed in the course formation of mutations was observed in each of the
of further culture maintenance. These were mutations following generations: lethal mutation 9 gave the
(1) cubitus interruptus, (2) radius incompletus, (3) black; plexus phenotype, which produced the speck pheno-
(4) dumpy, (5) flies with an abnormal tergite pattern, and type after several generations. In the next genera-
(6) flies with a bifurcated head; (7) flies with a flattened tion the latter gave the purple cinnabar and Bar
head and a long neck. phenotypes, which, in turn, produced upon the next
Dimorphic mutations. All mutations were initially transfer the phenotype alveolar eyes. Then followed
maintained in a female culture with attached-X chro- Dichaete, which gave rise to the phenotype broad
mosomes. After their transfer for being maintained in a wings, and then the phenotype raised wings fol-
heterozygote with a Muller-5 chromosome four muta- lowed (Fig. 6). All these phenotypic forms are
tions with unusual phenotypes were revealed: (1) short- maintained in special derivatives.

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 CONDITIONAL LETHAL MUTATIONS SHIFT THE GENOME 281

Table 3. The progeny of In(1)Muller-5, wa B/+ females carrying a mutation from crosses with yellow males*
Phenotypes of offsprings
Proportion
Male Total
females males of patroclinous y males
culture no. progeny
in the progeny
B + waB + y
1 65 5 56 1 47 174 0.27
2 93 59 80 31 54 317 0.17
3 28 19 32 23 20 122 0.16
5 52 63 60 53 14 242 0.06
6 147 105 102 74 73 501 0.15
7 124 86 64 68 35 377 0.09
8 83 88 72 89 31 363 0.09
9 88 44 37 31 50 250 0.20
10 50 49 51 44 0 194 0.00
11 76 66 72 63 14 291 0.05
27 54 38 36 21 49 198 0.25
29 77 85 44 60 19 285 0.07
30 109 70 85 58 56 378 0.15
31 132 89 83 77 79 460 0.17
32 57 65 57 44 3 226 0.01
33 45 31 53 23 10 162 0.06
34 99 63 42 27 33 264 0.13
35 142 56 85 37 124 444 0.28
36 138 97 98 100 48 481 0.10
38 140 107 119 120 18 504 0.04
41 93 88 66 84 28 359 0.08
Control 262 258 178 239 0 937 0.00
* The mutation was in the chromosome (+).

Single and Mass Modifications
Waves of phenocopies appeared in the mutation cul-
tures. One or another phenotype of the known mutations
was produced in one or several generations: black, purple,
brown, trident, abnormal abdomen, Notch, yellow, Dicha-
ete, etc. In most cases, several individuals of a new pheno-
type (from three to ten) appeared at once. As a rule, the
number of such individuals increased in the next genera-
tion. However, after several generations they gradually
disappeared from the progeny. Attempts to fix the new
phenotype in a derived strain failed. For example, one of
the frequent phenocopies were melanoma-like forma-
tions. Derivatives with 100% content of melanoma in an
offspring were sometimes obtained, but after some gener-
ations the phenotype was completely lost.
Small individuals periodically appeared in the mutation
cultures. Despite repeated attempts, no culture with such a
phenotype was obtained. Periodically, sharp deviations in
the sex ratio were observed. These mutants behaved as
described for phenocopies. Some modifications are pre-
sented in Fig. 7.
Mass Formation of Morphoses
As mentioned previously, various developmental
defects (morphoses) occurred in the mutation cultures and
in the progeny from crosses of mutants with other labora-
tory strains [8, 10–12]. The frequency of occurrence of mor-
phoses in cultures was far higher than that of secondary
mutations and modifications. The phenomenon of forma-
tion of morphoses constantly accompanied crosses of
mutants with other strains. In double heterozygotes for the
X chromosome, mutations 494 of the total progeny of 9795
individials (5.04%) had morphoses. In five cases the prog-
eny (>50 individuals) entirely consisted of flies with mor-
phoses. In the progeny of female heterozygotes for a muta-
tion in the X chromosome and in the marker X chromo-
some y2 ec cv ct v f, the proportion of offsprings with
morphoses varied from 2.4 to 26.0% (12.3% on the aver-
age) (Table 4).

RUSSIAN JOURNAL OF GENETICS Vol. 45 No. 3 2009

282 CHADOV et al.

(a) (b)

(c) (d)

(e) (f)

(g) (h)

Fig. 5. Formation of mosaics and gynandromorphs in cultures containing conditional lethals. (a) different colors of the eys in an
offspring of wa/+ female; (b) different forms of the eyes in an offspring of Ç/+ female; (c) the absence of coloration of the left half
of the last tergites; (d) the right half of the head and thorax is of the male type (a bar-shaped eye, genital eminence on the prothoracic
leg, yellow color of the legs), the remaining part of the body is of the female type; (e) the left half of the body is of the male type
(yellow color of the body, shortened wing), the right is female; the structure of the external genital organs is mixed; (f) the left half
of the abdomen is colored by the female type, the right one by the male type; (g) the left side of the individual has the male appear-
ance; (h) the right side of the same individual, including the external genital organs, has the female appearance.

RUSSIAN JOURNAL OF GENETICS Vol. 45 No. 3 2009

 CONDITIONAL LETHAL MUTATIONS SHIFT THE GENOME 283

(a) (b)

(c) (d)

(e) (f)

(g) (h)

Fig. 6. Formation of two complexes of mutations in cultures with conditional lethals. Complex (a)–(c): (a) phenotype interrupted
vein L2; (b) phenotypes bright red eyes and aristae without branching, control is above; (c) phenotype eye reduction, control is on
the right. Complex (d)–(h): (d) phenotype a dark spot at the base of the wing; (e) phenotypes purple, cinnabar, facet (eye facets are
large and orange); (f) phenotype broad wings; (g) phenotype parted wings; (h) phenotype parted and raised wings.

RUSSIAN JOURNAL OF GENETICS Vol. 45 No. 3 2009

284 CHADOV et al.

(a) (b)

(c) (d)

(e) (f)

(g) (h)

Fig. 7. Modifications in cultures containing conditional lethals. (a) melanoma, a tissue region of dark color; (b) wings with bubbles;
(c) pale dwarfs, small-size flies of pale yellow color; (d) vertically curved wings; (e) shortened wings, the wings are changed and
shortened up to their complete absence; (f) a dwarf, a male is smaller in size, normally colored, above is a normal male; (g) a dark-
colored female, above is a normally colored male; (h) three individuals with defects of the facet part of the eye.

RUSSIAN JOURNAL OF GENETICS Vol. 45 No. 3 2009

 CONDITIONAL LETHAL MUTATIONS SHIFT THE GENOME 285

Frequency of Occurrence of Modifications Table 4. Formation of morphoses in females “mutation/y2

and Secondary Mutations ec cv ct v f ”
The presented data on the formation of new pheno- Proportion of offsprings
Mutation no. Total progeny
types (mutations, modifications, and morphoses) are with morphoses, %
based on observations of mutation cultures in the
course of their maintenance or rarely on their use in 2 485 11.3
experiments. In cases when the facts were unexpected, 3 244 10.7
there were no strict estimations of the frequency. At the 5 362 26.0
same time, it can be stated that: (1) the frequencies of 6 596 2.4
the observed events are several orders of magnitude
higher than in cultures of ordinary Drosophila muta- 7 317 17.9
tions; (2) the process of formation of new phenotypes is 8 405 14.0
uneven, it is most intense immediately after a mutation 9 428 14.7
is obtained and in the case of its hybridization with 10 271 6.6
another culture; (3) the occurrence of new forms tends
to decrease in the course of maintaining a mutant in cul- 11 390 16.7
ture. 27 108 6.5
29 471 3.2
DISCUSSION 30 243 11.1
31 417 8.6
Observations of mutant cultures show that mutant
genomes are in the state of instability that has multiple 32 415 12.8
manifestations. Delethalization points to instability of 33 97 15.5
mutated loci themselves. Nonexpression of dominant 34 737 10.9
mutations in an opposite chromosome and the forma- 35 478 11.9
tion of new mutations indicate that instability is
extended to other loci. The loss and nondisjunction of 36 327 25.7
X chromosomes suggest disturbances in meiosis, and 38 126 3.2
the formation of mosaics and gynandromorphs is the 41 408 16.4
sign of mitotic disturbances. The abnormal character of
genetic processes in mitotically dividing cells is also Control 3687 0
demonstrated by the formation of morphoses. Morpho-
ses represent unilateral disturbances in the process of
ontogenesis in a limited group of somatic cells of a often considered independently. In general, genetic insta-
developing organism. bility is defined as a long-term preservation of deviations
in the organization and functioning of the genetic appara-
Instability concerns both structural genes (forma- tus, DNA-protein complex [14]. Instability manifests
tion of phenotypically known mutations) and regula- itself in the long-term formation with a high frequency of
tory genes controlling ontogenesis. The formation of point mutations, chromosome rearrangements, in carcino-
complex morphological structures in an improper place genesis, aneuploidy, and in chronic death of part of cells in
(morphoses) is explained by the initiation of a chain of cultures in vitro. The mechanisms of induction and main-
ontogenetic events automatically following one another tenance of instability are under discussion. In the case of
in cells that are not meant for that. irradiation, the cause of instability is considered to be
The facts of formation of phenotypically known direct damage of DNA and of the cytoplasm products.
mutations with stable inheritance over generations One of the causes of instability in carcinogenesis is
demonstrate that disturbances may be manifested as assumed to be aneuploidy [15] for which cis-regulatory
changes in the primary DNA sequence. However, dis- mechanisms of generation of instability in cell cultures
turbances may not involve the primary structure. Then were demonstrated [16]. The phenomenology of instabil-
the variant of a mutation for one generation only (a typ- ity is associated with the enhancement of transpositional
ical phenocopy) is realized or the variant of a transit activity of mobile genetic elements (MGEs). It is mobili-
mutation for several generations (waves of pheno- zation of retrovirus elements in the case of carcinogenesis
copies). In the latter case, a temporary change of a DNA and mobilization of P-M and I-R type MGEs in the case of
region in the generative pathway must take place, hybrid dysgenesis [16]. Irradiation is also an inducer of
which is transmitted to offsprings in a number of gen- transpositional activity of MGEs. Under the action of irra-
erations and then disappears. diation the rates of insertions increase more than by an
The phenomenon of genetic instability discovered by order of magnitude [17]. This work sheds new light on the
McClintock is widely spread. Instability is characteristic problem of instability. It can be inferred on the basis of the
of a number of phenomena, such as carcinogenesis, hybrid data obtained that the state of instability is a response of
dysgenesis, and late effects of radiation, although it is the genetic system to the damage of a special group of

RUSSIAN JOURNAL OF GENETICS Vol. 45 No. 3 2009

286 CHADOV et al.
genetic regions. These regions carry genes whose muta-
tions are expressed as conditional dominant lethals.
ACKNOWLEDGMENTS
The work was supported by the Russian Foundation for
Basic Research (grant nos. 04-04-48100 and 08-04-
00094a) and the Program of the Presidium of the Russian
Academy of Sciences “Origin and Evolution of the Bio-
sphere”.
REFERENCES
1. Chadov, B.F., Chadova, E.V., Kopyl, S.A., and
Fedorova, N.B., A New Class of Mutations in Drosophila
melanogaster, Dokl. Ross. Akad. Nauk, 2000, vol. 373, no. 5,
pp. 714–717.
2. Chadov, B.F., Mutations in the Regulatory Genes in Droso-
phila melanogaster, in Biodiversity and Dynamics of Ecosys-
tems in North Eurasia, Proc. Int. Conf., Novosibirsk, 2000,
pp. 16–18.
3. Chadov, B.F., Mutations Able to Initiate Speciation, Evolyut-
sionnaya biologiya (Evolutionary Biology), vol. 1, Stegnii,
V.N, Ed., Tomsk: Tomsk Gos. Univ., 2001, pp. 138–162.
4. Chadov, B.F., Chadova, E.V., Kopyl, S.A., et al., From Genet-
ics of Intraspecific Differences to Genetics of Intraspecific
Similarity, Russ. J. Genet., 2004, vol. 40, no. 9, pp. 1157–
1172.
5. Chadov, B.F., Attributes of Intraspecific Similarity and the
Specificity of Mendel’s Approach to Heredity, Filosofiya
Nauki, 2005, no. 3 (26), pp. 94–114.
6. Chadov, B.F., Chadova, E.V., Kopyl, S.A., and
Fedorova, N.B., Delayed Activation of the Maternal Genome
during Early Development of Drosophila, Dokl. Ross. Akad.
Nauk, 2001, vol. 378, no. 6, pp. 841–845.
7. Chadov, B.F., Chadova, E.V., Khotskina, E.A., et al., The
Main Effect of Chromosomal Rearrangement is Changing
the Action of Regulatory Genes, Russ. J. Genet., 2004,
vol. 40, no. 7, pp. 893–902.
RUSSIAN JOURNAL OF GENETICS
8. Chadov, B.F. and Fedorova, N.B., The Elementary
Event of Development, Dokl. Ross. Akad. Nauk,
2003, vol. 389, no. 3, pp. 408–412.
9. Chadov, B.F., The “Image” of the Regulatory Gene in
Experiments with Drosophila, Russ. J. Genet., 2002,
vol. 38, no. 7, pp. 869–880.
10. Chadov B.F. Facultative Dominant Lethals: Genetics,
Ontogeny, and Phylogeny, Evolyutsionnaya biologiya
(Evolutionary Biology), vol. 2, Stegnii, V.N., Ed.,
Tomsk: Tomsk Gos. Univ., 2001, pp. 118–142.
11. Chadov, B.F., Chadova, E.V., Kopyl, S.A., et al., Genes
Controlling Ontogeny: Morphosis, Phenocopies, Dimor-
phs, and other Visible Expressions of Mutant Genes,
Russ. J. Genet., 2004, vol. 40, no. 3, pp. 353–365.
12. Chadov, B.F., Ontogenes in Drosophila melanogaster:
Genetic Features and Role in Onto- and Phylogenesis,
Sovremennye problemy genetiki, radiobiologii, radi-
oekologii i evolyutsii (Modern Problems of Genetics,
Radiobiology, Radioecology, and Evolution), Proc. Sec-
ond Int. Conf. on the Occasion of the 105th Anniversary
of N.V. Timofeev-Resovsky), Korogodina, V.L,
Chin’i, A.A, and Durante, M., Eds., Dubna: Ob. Inst.
Yad. Issled., 2007, vol. 1, pp. 80–91.
13. Chadov, B.F., Chadova, E.V., Khotskina, E.A., and
Fedorova, N.B., Mutation in Ontogeny—Destabilizing
of Genome—Morphogenesis, Evolyutsionnaya
biologiya (Evolutionary Biology), vol. 3, Stegnii, V.N,
Ed., Tomsk: Tomsk Gos. Univ., 2001, pp. 92–106.
14. Lindsley, D.L. and Grell, E.H., Genetic Variation of
Drosophila melanogaster, Carnegie Inst. Wash. Publ.,
1968, no. 627, p. 472.
15. Bridges, C.B., Nondisjunction as Proof of the Chromo-
some Theory of Heredity, Genetics, 1916, vol. 1,
pp. 107–162.
16. Yarmonenko, S.P. and Vainson, A.A., Radiobiologiya
cheloveka i zhivotnykh (Human and Animal Radiobiol-
ogy), Moscow: Vysshaya Shkola, 2004.
17. Duesberg, P., Rausch, C., Rasnick, D., and
Hehlmann, R., Genetic Instability of Cancer Cells Is
Proportional to Their Degree of Aneuploidy, Proc. Natl.
Acad. Sci. USA, 1998, vol. 95, no. 23, pp. 13692–13697.
Vol. 45 No. 3 2009