Identification and characterization of a DNA primase activity present in herpes simplex virus type 1-infected HeLa cells

Journal Article · Tue Mar 01 00:00:00 EST 1988 · Journal of Virology; (USA)

OSTI ID:6796193

Holmes, A M; Wietstock, S M; Ruyechan, W T ^[1]

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1. Uniformed Services Univ. of the Health Sciences, Bethesda, MD (USA)

A novel DNA primase activity has been identified in HeLa cells infected with herpes simplex virus type 1 (HSV-1). Such an activity has not been detected in mock-infected cells. The primase activity coeluted with a portion of HSV-1 DNA polymerase from single-stranded DNA agarose columns loaded with high-salt extracts derived from infected cells. This DNA primase activity could be distinguished from host HeLa cell DNA primase by several criteria. First, the pH optimum of the HSV primase was relatively broad and peaked at 8.2 to 8.7 pH units. Second, freshly isolated HSV DNA primase was less salt sensitive than the HeLa primase. Third, antibodies raised against individual peptides of the calf thymus DNA polymerase:primase complex cross-reacted with the HeLa primase but did not react with the HSV DNA primase. Fourth, freshly prepared HSV DNA primase appeared to be associated with the HSV polymerase, but after storage at 4{degree}C for several weeks, the DNA primase separated from the viral DNA polymerase. This free DNA primase had an apparent molecular size of approximately 40 kilodaltons, whereas free HeLa DNA primase had an apparent molecular size of approximately 110 kilodaltons. On the basis of these data, the authors believe that the novel DNA primase activity in HSV-infected cells may be virus coded and that this enzyme represents a new and important function involved in the replication of HSV DNA.

• OSTI ID: 6796193
• Journal Information: Journal of Virology; (USA), Vol. 62:3; ISSN 0022-538X
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
DNA POLYMERASES
BIOCHEMISTRY
VIRUSES
ENZYME ACTIVITY
HELA CELLS
HERPES SIMPLEX
NUCLEOTIDES
PHOSPHORUS 32
TRITIUM COMPOUNDS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CHEMISTRY
DAYS LIVING RADIOISOTOPES
DISEASES
ENZYMES
HYDROGEN COMPOUNDS
INFECTIOUS DISEASES
ISOTOPES
LIGHT NUCLEI
MICROORGANISMS
NUCLEI
NUCLEOTIDYLTRANSFERASES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PARASITES
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
POLYMERASES
RADIOISOTOPES
SKIN DISEASES
TRANSFERASES
VIRAL DISEASES
550201* - Biochemistry- Tracer Techniques