DNA-containing phages neutralizing with anti-MS2 serum
The DNA-conlaining phages neutralizing with anti-MS2 serum have been detected in MS2-induced bacterial, cultures, in the cells containing the recombinant plasmid as well as in preparations of transducing lambda and PI phages. The identity of all these new developed bacteriophages permits to suppose...
Збережено в:
| Дата: | 1998 |
|---|---|
| Автор: | |
| Формат: | Стаття |
| Мова: | English |
| Опубліковано: |
Інститут молекулярної біології і генетики НАН України
1998
|
| Назва видання: | Биополимеры и клетка |
| Теми: | |
| Онлайн доступ: | http://dspace.nbuv.gov.ua/handle/123456789/157481 |
| Теги: |
Додати тег
Немає тегів, Будьте першим, хто поставить тег для цього запису!
|
| Назва журналу: | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| Цитувати: | DNA-containing phages neutralizing with anti-MS2 serum / T.P. Pererva // Биополимеры и клетка. — 1998. — Т. 14, № 6. — С. 549-552. — Бібліогр.: 11 назв. — англ. |
Репозитарії
Digital Library of Periodicals of National Academy of Sciences of Ukraine| id |
irk-123456789-157481 |
|---|---|
| record_format |
dspace |
| spelling |
irk-123456789-1574812019-07-04T22:00:16Z DNA-containing phages neutralizing with anti-MS2 serum Pererva, T.P. Вирусы и клетка The DNA-conlaining phages neutralizing with anti-MS2 serum have been detected in MS2-induced bacterial, cultures, in the cells containing the recombinant plasmid as well as in preparations of transducing lambda and PI phages. The identity of all these new developed bacteriophages permits to suppose the common mechanism of their origin. ДНК-вмісні фаги, які нейтралізуються анті-MS2 сироваткою, виявлено в культурах MS2-індукованих мутантів, у клітинах, що містять рекомбінантну плазміду, та в препаратах фагів MS2 і РІ, здатних до трансдукції фагів лямбда. Тотожність новоутворених бактеріофагів дозволяє припустити загальний механізм їхнього походження. ДНК-содержащие фаги, нейтрализующиеся анти-MS2 сывороткой, обнаружены з MS2-индуцированных бактериальных культурах, в клетках, содержащих рекомбинантную плазмиду, в препаратах фагов MS2, Р1 и лямбда-трансдуцирующих фагов. Идентичность новообразованных бактериофагов позволяет предположить общий механизм, их происхождения. 1998 Article DNA-containing phages neutralizing with anti-MS2 serum / T.P. Pererva // Биополимеры и клетка. — 1998. — Т. 14, № 6. — С. 549-552. — Бібліогр.: 11 назв. — англ. 0233-7657 DOI: http://dx.doi.org/10.7124/bc.0004F9 http://dspace.nbuv.gov.ua/handle/123456789/157481 en Биополимеры и клетка Інститут молекулярної біології і генетики НАН України |
| institution |
Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| collection |
DSpace DC |
| language |
English |
| topic |
Вирусы и клетка Вирусы и клетка |
| spellingShingle |
Вирусы и клетка Вирусы и клетка Pererva, T.P. DNA-containing phages neutralizing with anti-MS2 serum Биополимеры и клетка |
| description |
The DNA-conlaining phages neutralizing with anti-MS2 serum have been detected in MS2-induced bacterial, cultures, in the cells containing the recombinant plasmid as well as in preparations of transducing lambda and PI phages. The identity of all these new developed bacteriophages permits to suppose the common mechanism of their origin. |
| format |
Article |
| author |
Pererva, T.P. |
| author_facet |
Pererva, T.P. |
| author_sort |
Pererva, T.P. |
| title |
DNA-containing phages neutralizing with anti-MS2 serum |
| title_short |
DNA-containing phages neutralizing with anti-MS2 serum |
| title_full |
DNA-containing phages neutralizing with anti-MS2 serum |
| title_fullStr |
DNA-containing phages neutralizing with anti-MS2 serum |
| title_full_unstemmed |
DNA-containing phages neutralizing with anti-MS2 serum |
| title_sort |
dna-containing phages neutralizing with anti-ms2 serum |
| publisher |
Інститут молекулярної біології і генетики НАН України |
| publishDate |
1998 |
| topic_facet |
Вирусы и клетка |
| url |
http://dspace.nbuv.gov.ua/handle/123456789/157481 |
| citation_txt |
DNA-containing phages neutralizing with anti-MS2 serum / T.P. Pererva // Биополимеры и клетка. — 1998. — Т. 14, № 6. — С. 549-552. — Бібліогр.: 11 назв. — англ. |
| series |
Биополимеры и клетка |
| work_keys_str_mv |
AT perervatp dnacontainingphagesneutralizingwithantims2serum |
| first_indexed |
2025-07-14T08:24:32Z |
| last_indexed |
2025-07-14T08:24:32Z |
| _version_ |
1837610013339680768 |
| fulltext |
ISSN 0233-7657. Биополимеры и клетка. 1998. Т. 14. № 6
В И Р У С Ы И К Л Е Т К А
DN A-containing phages neutralizing
with anti-MS2 serum
Tamara P. Pererva
Institute of Molecular Biology and Genetics, National Academy of Sciences of Ukraine
150 Acad. Zabolotnogo str., Kyiv, 252143 , Ukraine
The DNA-containing phages neutralizing with anti-MS2 serum have been detected in MS2-inducco
bacterial, cultures, in the cells containing the recombinant plasmid as well as in preparations of transducing
lambda and PI phages. The identity of all these new developed bacteriophages permits to suppose the
common mechanism of their origin.
We have earlier described the induction of MS2-
resistant forms in the offspring of an Escherichia coli
AB 259 Hfr 3000 cell having survived this phage
infection 11 I. The phage-induced mutants have been
shown to be genetically unstable ones; they are able
to segregate new mutant types; two of them, a
granular mutant and a lysing one, have been studied
in detail [2 J. The cloning of the mutant region in a
non-replicating Ap-fragment [3] has demonstra ted
the some possibility of integration of an MS2-specific
sequence with the E, coli cell DNA; such a result
induces to study these mutants from the new point of
view of RNA phages-host cell interaction problem as
origin and properties of these mutants suggested the
presence at least of a part of phage genome or its
DNA copy in their chromosome. T h e at tempts to find
the association of MS2-specific nucleic acid with the
host cell chromosome has been made using the
method of hybridization of MS2-induced mutant RNA
and DNA with MS2 cDNA [4] and by the genetic
transfer of the mutant properties into the lambda
phage chromosome [5 ] . Despite of some positive
results we have failed to detect the presence of MS2
phage in MS2-induced mutan t cells or the phage
secretion by these cells. Such phenomena do not
correlate with the well-known tradit ional lysogeny
conception as well as with the persistant state con
ception because in both these cases bacterial cultures
contain some quantity of free phage. At the same
time, such fact as DNA-containing MS2-derivative
© T. p . P U R E R V A , 1998
phages segregation in the course of transducing PI
and lambda phages reproduction [5] suggest the
anti-MS2 serum neutral izing phages might have been
detected in MS2-induced mutan t cells of granular ana
lysing types. T h e aim of this work was revealing of
DNA-containing phages with some MS2-iikc pro-
perties and comparison of these properties with ones
of phage segregants described earlier [5 |.
Indeed a prolongated observation of wild type E.
coli 3000 cells put in the medium together with lysing
and granular type mutan ts permits to detect a phage
whose development goes with the extremely low-
frequency. If a drop of an one-day-old E. coli 3000
or of another F-pili forming culture is put on the lawn
formed by any of these two mutan t s a spot region of
the wild type lawn becomes markedly clearer in
several days . Some additional passages of the material
isolated from the spot give us the possibility to detect
the phages being identical and possessing the host
range of the MS2 phage. Contrary to phages ap
pearing in QB- and MS2-resistant fi + R E. con
cultures [6, 7 ] the phage particles found in our
test-system a re markedly different in their size and
morphology from the MS2 phage; these particles are
DNA-containing and consisted of heads and tails:
they have been neutral ized by the anti-MS2 serum,
the neutralization level being the same as with the
native MS2 phage. Neutral izat ion constants were 285,
267, 230 and 293 for MS2, phage segregated from
lysing bacterial mutant , phage segregated from granu
lar mutant and phage segregated from recombinant
plasmid-containing culture accordingly. Immunization
P H R J B R V A I . P
of rabbits was made according to Adams [8 J and
preliminary purifying of MS2 phage in CsCl gradient
was made according to Maniatis et al. [9 J. Anti
MS2-serum obtained in such a way did not neutralise
phages PI and A as well as it did not agglutinate E.
coli 3000 host cells. On the other side ant i -serums
against phages A and P I and to E. coli 3000 cells did
not neutralise both MS2 phage and all the new
DNA-contained phages obtained in the process of this
work. Exhaustion of anti MS2-serum by MS2 phage
preparations having different phage particles con
centrations decreased neutralizing properties of serum
both to MS2 phage and to DNA-containing phages
described here; in the cases when the high-t i t re M.S2
phage was used for exhaust ion neutralizing properties
of serum disappeared completely and all tested pha
ges (both RNA- and DNA-containing ones) formed
negative colonies in the presence of such exhausted
anti MS2-serum. It should be noted that these data
confirm only the activity of anti MS2-serum to new
phages but don ' t exlude the possibility of the pre
sence of some other ant igens on the surface of these
phages. This problem will be investigated by us in the
further experiments . It is clear from our electron
microscopic photoes of MS2 derivative phage particles
having been mixed with the T4 phage (Fig. 1) and
MS2 phage (Fig. 2) that the derivative particles are
markedly larger comparing to the MS2 ones. Their
icosahedrical head having a diameter about 50 nm,
and their tails being about 150—160 nm long. Despite
of these particles strict specificity for donor type cells,
our electron microscopic investigations detect no deri
vative particles adsorption on F-pili. The cause of this
phenomenon is supposed to be elucidated in further
investigations. At the same time such particles form
negative colonies belonging to the same type as the
colonies formed by P I and lambda transducing pha
ges segregants 15 J. Despite of the bacterial lawn
ageing, the colonies of DNA-containing MS2 d(
vative phages become larger during several days, the
demi-lysis zones becoming also wider and occuping
the whole lawn. The cells containing the recombinant
plasmid pL34 constructed by us [3] also accumulaic
the DNA-containing phage identical to those om
found in MS2-induced mutants from the point of view
of its morphology, its neutralization with anti-MS2
serum, and its restriction obtained after restriction
nucleases t reatment (see Fig. 3). The identical DNA
containing particles are also found in MS2 pre
parat ions, such an identity having been proved by the
markers already described above for other DNA
containing phages neutralizing with anti~-MS2 senmh
Fig. 1. Electron micro
graph of the DNA coniai
ning MS2-like phages mi
xod with 14 phage. Lira
nyl acetate (2 %> stai
ning. X 120000
550
L J N А - С О N Т А ININ G P H A G E S NEU 'i K A U / . I N ' G W I T H A N ! I MS 2 SKRI.'M
Fig. 3. Klectrophoregram of the. DNAs isolated from MS2--like phages
and digested with restnctascs Baelll (1—J), • PstI {4—6), and
H.imHJ (7—9) (1, 4, 7 - DNA of the phage segregated from E.
coli 3000 lysing type mutant; 2, 5, 8 — DNA of the phage
segregated from E. coli granular mutant; i , 6, 9 — DNA from the
phage segregated from pL34-containing cells); 10— lambda phage
DNA + licoRl + HindiU (marker lane)
T h e formation of DNA-contaming phages in
MS2-induced E. coli mutan t s suggests once more t h a i
some processes well known and investigated in deiah
may be accompanied with some other events of a verv
low frequency followed by new biological f o r m s b i r th
being new virus forms in our case. Contrary to
retrovirus-cell systems, no DNA-containing structures
taking part in RNA-containing phages reproduction
and phage RNA replication have been yet described.
Additionally, the construction of reveriase-target MS2
DNA in vitro is completely impossible without arti
ficial connection of the poly (dA) • poly (dT)-l inker to
MS2 phage RNA [10] being inaccessible for this
enzyme without such a supplement. So it cannot be
ruled out that the process of formation of DNA
containing phages antigenically related or identical to
RNA-containing ones may be due to other mecha
nisms of RNA-DNA interactions perhaps more similar
to those described previously for p r o c a r v o t i c a - a s
111], It cannot be ruled out that the presence in t h e
bacterial cell of RNA-phage specific information inde
pendently of its form (phage multipl lcation or some
other form specific for MS2-induced cell mutants)
induces prophages presented in E. coli. cells chromo
somes and plasmids. Such a prophage sequence m a y
form hybride forms with RNA phages components
both on the level of genomes and MS2-specific protein
coating of phage particles with new properties.
In any case, we have data enough to assume the
new virus-host cell interaction ideas are acceptable for
551
PJ3RERVA Т. p .
RNA-containing bacteriophages, at least at the struc
tural level, and may be successfully detected in a
system of MS2-induccd cellular and phagic mutants.
Acknowledgement. T h e au thor thanks Dr. F.
Tovkatch for the preparat ion of electron microscopic
photoes, Dr. Anna Miriuta for the restrictive analysis
of the phage DNAs, and Dr. Ella Zherebtsova having
prepared the ant i -phage serum and helped to prepare
this manuscript .
7. П. Перерва
ДНК-вмісні фаги, що нейтралізуються анти-М82 сироваткою
Резюме
ДНК-вмісні фаги, які нейтралізуються anmu~MS2 сироват
кою, виявлено в культурах MS2-indyкованих мутантів, у
клітинах, що містять рекомбінантну плазміду, та в препара
тах фагів MS2 і РІ, здатних до трансдукції фагів лямбда.
Тотожність новоутворених бактеріофагів дозволяє припусти
ти загальний механізм їхнього походження.
У, / / . Перерва
ДНК-содержащие бактериофаги, нейтрализующиеся airra-MS2
сывороткой
Резюме
ДНК-содержащие фаги, нейтрализующиеся anmu-MS2 сыво
роткой, обнаружены з MS2-индуцированных бактериальных
культурах, в клетках, содержащих рекомбинантную плазмиду,
в препаратах фагов MS2, Р1 и лямбда-трансдупирующих
фагов. Идентичность новообразованных бактериофагов позво
ляет предположить общий механизм, их происхождения.
REFERENCES
1. Перерва Т. 77. Устойчивость к фагу MS2, индуцированная
у Е. coli при заражении этим фагом / / Цитология и
генетика.—1977.—11 , № 1.—С. 3—9.
2. Перерва Т. П., Малюта С. С. Система MS2--индуци
рованных мутантов Е. coli по F-фактору / / Молекуляр.
биология.—Киев: Наук, думка, 1984.—Вып. 38. С. 81
90.
3. Перерва Т. П., Мирюта И. К)., Мир юта А. /О, Д ш о г е н и я
у фага MS2. Синтез фагоспецифичной РНК на клеточной
Д Н К / / Биополимеры и клетка. — 1 9 9 3 . — 9 , V_> 1.—С. 4 5 -
50.
4. Pererva Т. P., Miriuta N. Yu., Miriuta Л. Yu. el al. Analysis
of a recombinant plasmid containing MS2-iike sequence /7
Ib id .—1995.—11, N l . — P . 6 1 - 6 5 .
З.Перерва Т. П., Мирюта А. Ю., Вудмаска Л/. //., Аяек-
сеенко И. 77. Лизогения у фага MS2. Экспрессия M S 2 -
специфической информации сегрегантами нестабильны*
трансдуцирующих фагов Р1 и лямбда / / Гам же.—1996.—-
12, № 4 .—С. 7 3 — 8 3 .
6. Widmer И. Я, Lebec G. Der Einfluss von R \ S - P ! m g e n auf
Konjugationsfaktoren / / Pathol. Microbiol.--1(>74.•--•-••40, N 3
4.—S. 153—154.
7. Widmer H. Я, Lebec G. Wietere Ergcbnisse zor Wecnsel
wirkung zwischen RNS-Phagen und R-Fakloren / / ibid. 41
N 3 — 4 . — S . 194—195.
8. Adams M. Bacteriophages.—New York; London: im. Publ.
Inc., 1951.
9. Mauuamuc Т., Фрич Э., Сэмбрук Дж. Молекулярное кло
нирование—М.: Мир, 1984.
10. Devos Я, Van Emmelo J., Contreras Я, Tiers W. Construc
tion and characterization of a plasmid containing a i i c n h
full-size DNA copy of bacteriophage M S 2 RNA .// J. Мої
Biol .—1979.—N 4.—P. 5 9 5 — 6 1 9 .
\l. Inouye S.} Jnouye M. Bacterial reverse transeripiase 7 {•••
verse Transcriptase / / Eds A. M. Skalka, S. P. Gos l - \ i
York: Cold Spring Harb. Lab. press, 1 9 9 3 . - 492 p.
R e c e i v e d 2 4 . Ї 0 . ^ 7
552
|