The mode of replication of vaccinia virus DNA

Beatriz G.T. Pogo; Michael T. O'Shea

doi:10.1016/0042-6822(78)90213-1

The mode of replication of vaccinia virus DNA

Beatriz G.T. Pogo, Michael T. O'Shea

Research output: Contribution to journal › Article › peer-review

19 Scopus citations

Abstract

Synthesis of vaccinia virus DNA was studied by following the incorporation of [³H]thymidine in the cytoplasm of infected cells. During short pulses (1-5 min) the radioactive precursor was incorporated into single-stranded DNA fragments that sedimented as 10-12 S in neutral sucrose gradients. After the chase, these fragments were converted into a 34 S intermediate before becoming mature size molecules (68 S). Following longer periods of chase, the labeled DNA sedimented as cross-linked molecules in alkaline sucrose gradients (91-106 S) suggesting that the formation of cross-links is a postreplicative process. The presence of RNA covalently linked to DNA in the 12 S fragments was suggested by a shift in the equilibrium density sedimentation and by sensitivity to spleen phosphodiesterase after alkaline hydrolysis. Determination of the molecular weight of vaccinia DNA synthesized in cultures containing bromodeoxyuridine (BrUdR) demonstrated that replication proceeds in two directions.

Original language	English
Pages (from-to)	1-8
Number of pages	8
Journal	Virology
Volume	84
Issue number	1
DOIs	https://doi.org/10.1016/0042-6822(78)90213-1
State	Published - Jan 1978
Externally published	Yes

Access to Document

10.1016/0042-6822(78)90213-1

Cite this

@article{329036933e7744dcba2e470d67699a64,

title = "The mode of replication of vaccinia virus DNA",

abstract = "Synthesis of vaccinia virus DNA was studied by following the incorporation of [3H]thymidine in the cytoplasm of infected cells. During short pulses (1-5 min) the radioactive precursor was incorporated into single-stranded DNA fragments that sedimented as 10-12 S in neutral sucrose gradients. After the chase, these fragments were converted into a 34 S intermediate before becoming mature size molecules (68 S). Following longer periods of chase, the labeled DNA sedimented as cross-linked molecules in alkaline sucrose gradients (91-106 S) suggesting that the formation of cross-links is a postreplicative process. The presence of RNA covalently linked to DNA in the 12 S fragments was suggested by a shift in the equilibrium density sedimentation and by sensitivity to spleen phosphodiesterase after alkaline hydrolysis. Determination of the molecular weight of vaccinia DNA synthesized in cultures containing bromodeoxyuridine (BrUdR) demonstrated that replication proceeds in two directions.",

author = "Pogo, {Beatriz G.T.} and O'Shea, {Michael T.}",

note = "Funding Information: These studies were supported in part by Grant No. Al-12188 from the National Institute of Allergy and Infectious Diseases, U.S. Public Health Service.",

year = "1978",

month = jan,

doi = "10.1016/0042-6822(78)90213-1",

language = "English",

volume = "84",

pages = "1--8",

journal = "Virology",

issn = "0042-6822",

publisher = "Academic Press Inc.",

number = "1",

}

TY - JOUR

T1 - The mode of replication of vaccinia virus DNA

AU - Pogo, Beatriz G.T.

AU - O'Shea, Michael T.

N1 - Funding Information: These studies were supported in part by Grant No. Al-12188 from the National Institute of Allergy and Infectious Diseases, U.S. Public Health Service.

PY - 1978/1

Y1 - 1978/1

N2 - Synthesis of vaccinia virus DNA was studied by following the incorporation of [3H]thymidine in the cytoplasm of infected cells. During short pulses (1-5 min) the radioactive precursor was incorporated into single-stranded DNA fragments that sedimented as 10-12 S in neutral sucrose gradients. After the chase, these fragments were converted into a 34 S intermediate before becoming mature size molecules (68 S). Following longer periods of chase, the labeled DNA sedimented as cross-linked molecules in alkaline sucrose gradients (91-106 S) suggesting that the formation of cross-links is a postreplicative process. The presence of RNA covalently linked to DNA in the 12 S fragments was suggested by a shift in the equilibrium density sedimentation and by sensitivity to spleen phosphodiesterase after alkaline hydrolysis. Determination of the molecular weight of vaccinia DNA synthesized in cultures containing bromodeoxyuridine (BrUdR) demonstrated that replication proceeds in two directions.

AB - Synthesis of vaccinia virus DNA was studied by following the incorporation of [3H]thymidine in the cytoplasm of infected cells. During short pulses (1-5 min) the radioactive precursor was incorporated into single-stranded DNA fragments that sedimented as 10-12 S in neutral sucrose gradients. After the chase, these fragments were converted into a 34 S intermediate before becoming mature size molecules (68 S). Following longer periods of chase, the labeled DNA sedimented as cross-linked molecules in alkaline sucrose gradients (91-106 S) suggesting that the formation of cross-links is a postreplicative process. The presence of RNA covalently linked to DNA in the 12 S fragments was suggested by a shift in the equilibrium density sedimentation and by sensitivity to spleen phosphodiesterase after alkaline hydrolysis. Determination of the molecular weight of vaccinia DNA synthesized in cultures containing bromodeoxyuridine (BrUdR) demonstrated that replication proceeds in two directions.

UR - http://www.scopus.com/inward/record.url?scp=0017801177&partnerID=8YFLogxK

U2 - 10.1016/0042-6822(78)90213-1

DO - 10.1016/0042-6822(78)90213-1

M3 - Article

C2 - 619484

AN - SCOPUS:0017801177

SN - 0042-6822

VL - 84

SP - 1

EP - 8

JO - Virology

JF - Virology

IS - 1

ER -

The mode of replication of vaccinia virus DNA

Abstract

Access to Document

Fingerprint

Cite this