High selectivity of mutagenic action of synthetic polynucleotides
Solutions of high molecular weight syntetic poiyribo- and polydeoxyribonucleotides of known chemical structure injected into Drosophila melanogaster males caused a 6–17 fold increase of the overall frequency of the recessive lethals in the 2nd chromosome. Complementation analysis of the induced leth...
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Інститут молекулярної біології і генетики НАН України
1999
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irk-123456789-1565352019-07-05T17:11:08Z High selectivity of mutagenic action of synthetic polynucleotides Alexandrov, Yu.N. Обзоры Solutions of high molecular weight syntetic poiyribo- and polydeoxyribonucleotides of known chemical structure injected into Drosophila melanogaster males caused a 6–17 fold increase of the overall frequency of the recessive lethals in the 2nd chromosome. Complementation analysis of the induced lethals showed an extremely high locus-specificity for some of these mutagens. Possible mechanisms of the mutagenic action of synthetic polynucleotides are discussed. Високомолекулярні розчини полірибо- та полідезоксирибонуклеотидів, шр введено у самців Drosophila melanogaster, збільшували у 6–17 разів частоту рецесивних летальнних мутацій у 2-й хромосомі. Комплементаційний аналіз індукованих летальних мутацій виявив високу локус-специфічність для цих мутагенів. Обговорюються можливі механізми мутагенної дії синтетичних полінуклеотидів. Растворы полирибо- и полидезоксирибонуклеотидов известной химической структуры и высокой молекулярной массы, инъецированные в самцов Drosophila melanogaster, повышали в 6–17 раз частоту рецессивных летальных мутаций во 2-й хромосоме. Комплементационный анализ индуцированных летальных мутаций обнаружил высокую локус-специфичность для этих мутагенов. Обсуждаются возможные механизмы мутагенного действия синтетических полинуклеотидов. 1999 Article High selectivity of mutagenic action of synthetic polynucleotides / Yu.N. Alexandrov // Биополимеры и клетка. — 1999. — Т. 15, № 4. — С. 257-261. — Бібліогр.: 19 назв. — англ. 0233-7657 DOI: http://dx.doi.org/10.7124/bc.000522 http://dspace.nbuv.gov.ua/handle/123456789/156535 577.1 en Биополимеры и клетка Інститут молекулярної біології і генетики НАН України |
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Обзоры Обзоры Alexandrov, Yu.N. High selectivity of mutagenic action of synthetic polynucleotides Биополимеры и клетка |
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Solutions of high molecular weight syntetic poiyribo- and polydeoxyribonucleotides of known chemical structure injected into Drosophila melanogaster males caused a 6–17 fold increase of the overall frequency of the recessive lethals in the 2nd chromosome. Complementation analysis of the induced lethals showed an extremely high locus-specificity for some of these mutagens. Possible mechanisms of the mutagenic action of synthetic polynucleotides are discussed. |
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Alexandrov, Yu.N. |
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Alexandrov, Yu.N. |
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Alexandrov, Yu.N. |
| title |
High selectivity of mutagenic action of synthetic polynucleotides |
| title_short |
High selectivity of mutagenic action of synthetic polynucleotides |
| title_full |
High selectivity of mutagenic action of synthetic polynucleotides |
| title_fullStr |
High selectivity of mutagenic action of synthetic polynucleotides |
| title_full_unstemmed |
High selectivity of mutagenic action of synthetic polynucleotides |
| title_sort |
high selectivity of mutagenic action of synthetic polynucleotides |
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Інститут молекулярної біології і генетики НАН України |
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1999 |
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http://dspace.nbuv.gov.ua/handle/123456789/156535 |
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High selectivity of mutagenic action of synthetic polynucleotides / Yu.N. Alexandrov // Биополимеры и клетка. — 1999. — Т. 15, № 4. — С. 257-261. — Бібліогр.: 19 назв. — англ. |
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Биополимеры и клетка |
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AT alexandrovyun highselectivityofmutagenicactionofsyntheticpolynucleotides |
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1837611768372789248 |
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ISSN 0233-7657. Биополимеры и клетка. 1999. Т. 15. № 4
ОБЗОРЫ
High selectivity of mutagenic action of synthetic
polynucleotides
Yu. N. Alexandrov
Institute of Plant Physiology and Genetics National Academy of Sciences of Ukraine
148 Acad. Zabolotnoho vul., Kyiv, 03143 , Ukraine
Solutions of high molecular weight syntetic polyribo- and polydeoxyribonucleotid.es of known chemical
structure injected into Drosophila melanogaster males caused a 6—17 fold increase of the overall frequency
of the recessive lethals in the 2nd chromosome. Complementation analysis of the induced lethals showed
an extremely high locus-specificity for some of these mutagens. Possible mechanisms of the mutagenic
action of synthetic polynucleotides are discussed.
The fact that DNA isolated from various organisms
and from DNA-containing viruses is mutagenic for
Drosophila and shows a high specificity, inducing
mutations preferentially in certain chromosome loci
[1, 21 led to the supposition that synthetic poly
nucleotides of sufficient length likewise possess such
peculiar mutagenic properties. It seemed possible that
this could be true not only for polydeoxynucleotides
but also for polyribonucleotides. Some data on the
mutagenicity of natural RNA have been previously
obtained by Fahmy and Fahmy [3 ] , and in our
laboratory it had been shown that RNA-containing
viruses are mutagenic for Drosophila and that the
mutagenicity of these viruses is highly locus-specific
[1] . Moreover, our data led to the conclusion that
mutagenicity of viruses schould be probably attributed
solely to their nucleic acid and not their proteins.
Here we describe the results of experiments on the
induction of mutations in Drosophila by ten synthetic
polynucleotides of known chemical structure. Fahmy
and Fahmy [4] described the mutagenic action for
Drosophila of some synthetic polynucleotides but their
data are not very convincing as these data are based
chiefly on the frequency of Minute mutations which
are but weakly expressed so that it is extremely
difficult to discern them from wild type.
We tested the mutagenic action the following
polynucleotides: three of them, poly (dA), poly(dT)
and poly(dA, dT) were polydeoxyribonucleotides;
© Yu. N. A L E X A N D R O V , 1999
and seven, poly (A), poly (I), poly(U), poly (A, U) ,
poly (A, C), poly (A, I) poly (A, C, U) were polyri
bonucleotides. All these preparations consisted of a
mixture of molecules with a weight from 5 - Ю 5 to
16.5-10 6 Da. Poly(A), poly(A, C), poly(A, C, U)
were manufactured by «Serva» (Germany); poly(U)
by «Fluka» (Switzerland); poly(dA, dT) by «Bioche-
mical Inc.» (USA); all other polynucleotides were
synthetized in the Institute of Cytology and Genetics
(Rossia). Solutions of these preparations in saline
(0.9 % NaCl) were injected into young adult Dro
sophila melanogaster males of a wild-type stock D18
standartly used in our laboratory, each male received
ca. 0.25 jug of one of the polynucleotides dissolved in
saline. The males were previously proved to be free
from 2nd chromosome recessive lethals which could
have accumulated in the laboratory stock. This was
done by making the 2nd chromosome homozygous
(using balancers) just before the beginning of our
experiments. A search for recessive lethals induced by
polynucleotides in the 2nd chromosome of treated
males was carried out by the routine C l / L 2 method.
All the lethal-carrying chromosomes were produced
by different males, this excluding the possibility of
«clustering», i. e. an appearance of several identical
lethals as a result of mitotic proliferation of a gonial
chromosome. Every lethal was further genetically
verified. Control males received injections of saline.
Injection of polynucleotides induced recessive
lethals in the 2nd chromosome with a much higher
frequency then in the control as shown in Table 1.
257
http://polydeoxyribonucleotid.es
A L E X A N D R O V Yu. N.
Table 1
Frequency of recessive lethals induced by polynucleotides in the second chromosome
The number of loci in the 2nd chromosome
capable of mutating to a recessive lethal condition,
according to recent data [5, 6 ] , is about 1400—2000.
Our data also show that the number of such loci is
well over 1000. In contradistinction to this, the
number of loci in the 2nd chromosome where reces
sive lethal mutations were induced by polynucleotides
is very much less but in each of these loci mutations
arose unusually frequently, just as seen in case of
mutations caused by exogenous DNAs [1, 2 ] . This
was established by complementation tests aimed at
the determination of the frequency of allelism among
the lethals induced by each of the polynucleotides
(Table 2).
The minimal number of loci capable of mutating
to recessive lethality is inverse to the probability that
one such lethal is allelic to another. On the basic of
these calculations we can roughly determine the
number of loci in the 2nd chromosome which can
mutate to recessive lethal condition under the influ
ence of polynucleotides tested. The results are pre
sented in Table 3. For comparison, it is assumed that
in the 2nd chromosome there exist some 1600 loci
which can mutate to recessive lethality (this number
seems to be a fairly good guess).
This method of calculations gives only the lowest
number of loci. Their actual number may be some
what higher because of differencies in mutation rate
of different genes and the possible presence of
overlapping deletions possessing a recessive lethal
effect. But this qualification cannot blur the very great
difference between the number of loci which can
mutate spontaneously or under the influence of con
ventional mutagens and the number of loci affected by
the polynucleotides tested. The specificity of the
mutagenic action of these polynucleotides closely
resembles that of the mutagenic action of exogenous
DNAs but here it is even more pronounced. Nothing
comparable with this extraordinary locus-specificity
exists in the action of known physical or chemical
mutagens and none of then cause such an enormous
increase of the mutation rate of individual loci.
The high specificity of the mutagenic action of
synthetic polynucleotides is further confirmed by the
peculiar pattern of allelism among mutations caused
by them. This pattern is similar to that observed in
case of mutations induced by exogenous DNAs. Many
lethals are allelic to others which are not allelic to
each other. Complementation analysis and mapping of
lethals induced by polynucleotides show that this can
be attributed to two causes. It seems that rather often
several different lethals arise simultaneously in the
same chromosome of treated male. In other cases it is
possible that the polynucleotides affect several adje-
cent subunits (sites, neighbouring genes or groups of
genes) which can mutate separately or together.
Existence of chromosomes with multiple mutations
has been proved by the recombination analysis.
The spectra of induced lethal mutations differs
for different polynucleotides but this difference is not
258
HIGH SELECTIVITY O F M U T A G E N I C A C T I O N OF P O L Y N U C L E O T I D E S
*Both figures are probably considerably exaggerated. Ives studies lethals found in wild populations where identical recessive mutations are
usually frequent bacause of gene draft, and Wallace worked with lethals induced by radiation causing numerous deletions, many of which
may include several loci.
Table 3
Approximate frequency per locus of recessive lethals induced by some polynucleotides in the second chromosome
absolute. Some cases were found when the same lethal
was induced in the same locus by two different
polynucleotides.
Microscopic investigation of salyvary gland chro
mosomes and tests on dominant lethals showed that
the polynucleotides in our experiments do not induce
gross chromosome rearrangements. Evidently, all the
recessive lethals found were gene mutations or micro
scopically indiscernible microdeletions.
At present only speculative explanations may be
proposed for the mechanism of mutagenic action of
synthetic polynucleotides. Clearly, there exists a close
similarity in the action of these mutagens and the
mutagenic action of exogenous DNAs. This resem-
259
A L E X A N D R O V Yu. N.
blance is expressed in the very high frequency of gene
mutations or microdeletions preferentially induced in
a restricted number of chromosome loci; also, in the
non-random distribution of mutations along the chro
mosome and in the absence of gross chromosome
rearrangements. A number of considerations led to
the supposition that fragments of a foreing DNA
become inserted into the chromosome of the recipient
but only in such loci to which fragments of a given
DNA have some kind of affinity. This view is
supported by the fact that DNA of Drosophila itself is
also mutagenic for this insect but completely lacks
locus-specificity, probably because fragments of Dro
sophila DNA are homologous to a great number of loci
in the DNA of affected chromosomes [1 ].
The mutagenic action of synthetic polynucleotides
studied by us (its high locus-specificity, frequent
induction of multimutational events) closely resembles
that of exogenous DNAs, this suggesting a common
mechanism. However, the hypothetical insertion mo
del proposed by us needs serious amendments to
become applicable not only to synthetic polynuc
leotides of the desoxyribose type but also to the
ribose type. This amendment is also needed to
explain the mutagenic action of RNA-containing viru
ses as our experiments showed that their mutagenicity
is caused by their RNA (viral proteins complety
lacking mutagenic properties). But on theoretical
grounds it is absolutely impossible for fragments of
ribose polynucleotides or of natural RNAs to be
inserted into the chromosomal DNA of recipients. To
overcome this difficulty we supposed that not frag
ments of the molecules of the ribose type are inserted
into the chromosomal DNA but their DNA-copies
(cDNA) synthetized by reverse transcription in the
cells of the recipient; the enzymes necessary for
reverse transcription are present in insect cells [9,
10] .
Some doubts about the correctness of this exp
lanation arose after we showed that the spectrum of
lethal mutations induced in the 2nd chromosome of
D. melanogaster by injection of a solution of synthetic
poly(dA) is, against our expectation, suite different
from the spectrum of such mutations induced by
poly(dT) and that no correlation exist also between
the spectra of lethals induced by poly(U) and po
ly (dT). But eventually we realized that these facts
may not contradict our supposition that the mutagenic
action of synthetic polynucleotides of the ribose type
depends on the formation of their DNA copies by
reverse transcription. It is quite possible that an
insertion into a gene (or close to it) of a fragment of
poly(dA) will change its fine structure quite dif
ferently compare to insertion of poly(dT) etc.
A second possible mechanism of the mutagenic
action of exogenous DNAs and synthetic polynuc
leotides should also be mentioned. Alexandrov and
Gershenson [11 ] and Alexandrov [12] supposed that
these mutagenes can somehow activate mobile genetic
elements present in the Drosophila genome, this
leading to their transpositions causing mutations in
target loci. In favour of this idea, though indirectly,
are the numerous published data on the mutagenicity
of mobile genetic elements and especially the results
of Gerasimova and her collaborators [13—18] sho
wing that spontaneous multimutational events studied
by them in Drosophila are connected with and
seemingly caused by simultaneously occuring trans
positions of several different mobile genetic elements.
The possible mechanisms discussed above of the
mutagenic action of exogenous DNAs and synthetic
polynucleotides are not necessarily mutually exclusive
and may be complementary. Of course, we fully
realise that they are only speculative and their
verification, refutation or modification must await an
accumulation of additional experimental data. Until
then it is perhaps safer to limit ourselves to a vague
statement that, perephrasing Green [19] , exogenous
DNAs and synthetic polynucleotides act as inducers
of genomic stress, thereby increasing the occurrence
of gene mutations.
Acknowledgements. I am highly indebted to
S. M. Gershenson for his interest in this work, ever-
ready help, useful suggestions and critical reading of
the manuscript.
Ю. M. Александров
Висока селективність мутагеної дії синтетичних полінуклеотидів
Резюме
Високомолекулярні розчини полірибо- та полідезоксирибонук-
леотидів, шр введено у самців Drosophila melanogaster, збіль
шували у 6—17 разів частоту рецесивних летальнних мутацій
у 2-й хромосомі. Комплементаційний аналіз індукованих ле
тальних мутацій виявив високу локус-специфічність для цих
мутагенів. Обговорюються можливі механізми мутагенної дії
синтетичних полінуклеотидів.
Ю. Н. Александров
Высокая селективность мутагенного действия синтетических
полинуклеотидов
Резюме
Растворы полирибо- и полидезоксирибонуклеотидов известной
химической структуры и высокой молекулярной массы, инъе
цированные в самцов Drosophila melanogaster, повышали в 6—17
раз частоту рецессивных летальных мутаций во 2-й хромосо
ме. Комплементационный анализ индуцированных летальных
мутаций обнаружил высокую локус-специфичность для этих
мутагенов. Обсуждаются возможные механизмы мутагенного
действия синтетических полинуклеотидов.
260
HIGH SELECTIVITY O F M U T A G E N I C ACTION OF P O L Y N U C L E O T I D E S
REFERENCES
1. Gershenson S. M., Alexatidrov Yu. N.t Maliuta S. S.
Mutagenic action of DNA and viruses in Drosophila.—Kiyv:
Naukova dumka, 1975.—160 pp. (in Russian a. English).
2. Gershenson S. M. Mutagenic action of DNA, insertion, trans
position and gene instability / / Proc. 14 Int. Congr. of
Genet—Moscow: Mir, 1980.—Vol. 1, book 2 .—P. 91 — 115.
3. Fahmy О. C , Fahmy M. J. Mutagenic activity of macro-
molecules in Drosophila melanogaster II Nature.—1962.—
196, N 4857 .—P. 873—876 .
4. Fahmy O. G., Fahmy M. J. Mutagenesis in relation to nucleic
acid synthesis in cellular genetic system / / Mechanisms of
mutation and inducing factors.—Prague: Academia, 1966.—
P. 267—282 .
5. Simmons M. J., Crow J. F. Mutations affecting fitness in
Drosophila populations / / Ann. Rev. Genet .—1977 .—11 .—
P. 49—78.
6. Abrahamson S.y Wurgler E. F., De Jong C, Meyer H. V. How
many loci on the X-chromosome of Drosophila melanogaster
can mutate to recessive lethals? / / Envirom. Mutagens.—
1980 .—2.—P. 447—453 .
7. Ives P. T. The genetic structure of American populations of
Drosophila melanogaster II Genet ics .—1945.—30.—P. 167—
196.
8. Wallace B. Allelism of second chromosome lethals in Dro
sophila melanogaster II Proc. Nat. Acad. Sci USA.—1950 .—
36, N 11.—P. 645—657 .
9. Cohen L Penner P. E.t Loeb L A. Multiple DNA
polymerases displayed by isoelectric focusing / / Ann. N. Y.
Acad. Sc i .—1973 .—209, N 1.—P. 354—362 .
10. Margulies L, Chargaff E. Survey of DNA polymerase activity
during the early development of Drosophila melanogaster II
Proc. Nat. Acad. Sci USA.—1973 .—70 , N 10.—P. 2946—
2950.
11. Alexandrov Yu., Gershenson S. M. Mutagenic action of natural
and synthetic polynucleotides and the problem of directed
mutations / / Zhurn. Obsh. Bio l .—1982.—43, N 6.—P. 747—
763 (in Russian).
12. Alexandrov Yu. N. Natural and synthetic polynucleotides of
selective action / / Mutagenic action of natural and synthetic
polynucleotides / Ed. V. V. Morgun.—Kiev: Nauk. Dumka,
1988.—P. 4—10 (in Russian).
13. Gerasimova Т. I., Ilyin Yu. V., Misrokhi L. I., Semenova L.
V., Georgiev G. P. A direct demonstration of transposition of a
mobile element and its role in the appearance of unstable
mutations in Drosophila II Dokl. Akad. Nauk SSSR.—1983 .—
271 .—P. 977—980 (in Russian).
14. Gerasimova T. /., Mizrokhi L. Yu., Georgiev G. P. «Transposi-
tion bursts* in individual germ cells during genetic destabiliza-
tion in Drosophila melanogaster II Dokl. Akad. Nauk SSSR —
1984.—274, N 6.—P 1473—1476 (in Russian).
15. Gerasimova Т. I. Transposition of mobile elements and their
role in insertion mutagenesis in Drosophila II Stability and
variability of the genome.—Moscow: Nauka, 1985.—P. 26—39
(in Russian)
16. Gerasimova T. /., Mizrokhi L. Yu., Obolenkova L. A.,
Georgiev G. P. Directed mutagenesis under the influence of
mobile elements in unstable strains of Drosophila melanogaster
II Dokl. Akad. Nauk S S S R . — 1 9 8 5 . — 2 8 0 . — P . 1246—1250
(in Russian).
17. Gerasimova Т. I., Mitin V. A., Tchuricov N. A., Kochieva E.
Z. Site-specific mutagenesis in the cut locus of Drosophila
melanogaster under the influence of mobile element mdg4 II
Dokl. Akad. Nauk SSSR.—1988 .—300 .—P. 4 6 2 — 4 6 4 (in
Russian).
18. Gerasimova Т. I., Ladvishenko А. В., Mogila V. A., Georgieva
S. G.y Kiseleva S. M., Maxymiv D. V. Transposition bursts and
chromosome rearrangements in unstable Drosophila strains / /
Genet ika.—1990.—26.—P. 391—411 (in Russian).
19. Green M. M. Mobile elements and spontaneous gene mutations
/ / Bambury Rep. 30: Eukaryotic transposable elements as
mutagenic agents.—New York: Cold Spring Harbor Lab.,
1988.—P. 4 1 — 5 0 .
УДК 577.1
Received 28.04.98
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