Abstract

The objective of the research is to define possible controls for DNA biosynthesis during normal and abnormal growth. Most of the DNA polymerase Alpha activity from HeLa cells in S-phase has been isolated as a Mr 640,000 multiprotein form that is designated DNA polymerase Alpha2. The enzyme has been purified to electrophoretic homogeneity and resolved into its components that include; a Mr 180,000 polymerase Alpha subunit, a Mr 70,000 DNA primase, a Mr 69,000 3'---greater than 5' exonuclease, Mr 96,000 C1 and Mr 52,000 C2 primer-recognition proteins and Mr 92,000 Ap A binding protein. The interaction of DNA polymerase Alpha2 and other required proteins for the efficient and specific initiation of RNA primer synthesis will be defined. The conditions and protein requirements in addition to polymerase Alpha2 that are required for elongation of DNA chains with catalytic efficiency, processivity and fidelity will be ascertained. The enzymic and nonenzymic proteins in addition to DNA polymerase Alpha2 that are required for the replicationof SV40-DNA in vitro in the presence of T-antigen will be studied.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
2R01CA015187-11A1
Application #
3164114
Study Section
Biochemistry Study Section (BIO)
Project Start
1978-04-01
Project End
1991-11-30
Budget Start
1986-12-01
Budget End
1987-11-30
Support Year
11
Fiscal Year
1987
Total Cost
Indirect Cost

  Institution

Name
Worcester Foundation for Biomedical Research
Department
Type
DUNS #
City
Shrewsbury
State
MA
Country
United States
Zip Code
01545

  Publications

Göke, Friederike; Franzen, Alina; Hinz, Trista K et al. (2015) FGFR1 Expression Levels Predict BGJ398 Sensitivity of FGFR1-Dependent Head and Neck Squamous Cell Cancers. Clin Cancer Res 21:4356-64
Cao, Q P; McGrath, C A; Baril, E F et al. (1995) The 68 kDa calmodulin-binding protein is tightly associated with the multiprotein DNA polymerase alpha-primase complex in HeLa cells. Biochemistry 34:3878-83
Li, C; Goodchild, J; Baril, E F (1994) DNA ligase I is associated with the 21 S complex of enzymes for DNA synthesis in HeLa cells. Nucleic Acids Res 22:632-8
Cao, Q P; Pitt, S; Leszyk, J et al. (1994) DNA-dependent ATPase from HeLa cells is related to human Ku autoantigen. Biochemistry 33:8548-57
Li, C; Cao, L G; Wang, Y L et al. (1993) Further purification and characterization of a multienzyme complex for DNA synthesis in human cells. J Cell Biochem 53:405-19
Goodchild, J; Vishwanatha, J K (1991) A novel assay for DNA ligase. Nucleic Acids Res 19:3745
Vishwanatha, J K; Baril, E F (1990) Single-stranded-DNA-dependent ATPase from HeLa cells that stimulates DNA polymerase alpha-primase activity: purification and characterization of the ATPase. Biochemistry 29:8753-9
Malkas, L H; Hickey, R J; Li, C et al. (1990) A 21S enzyme complex from HeLa cells that functions in simian virus 40 DNA replication in vitro. Biochemistry 29:6362-74
Malkas, L H; Baril, E F (1989) Sequence recognition protein for the 17-base-pair A + T-rich tract in the replication origin of simian virus 40 DNA. Proc Natl Acad Sci U S A 86:70-4
SenGupta, D N; Kumar, P; Zmudzka, B Z et al. (1987) Mammalian alpha-polymerase: cloning of partial complementary DNA and immunobinding of catalytic subunit in crude homogenate protein blots. Biochemistry 26:956-63

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