Cloning and expression of cDNA for human poly(ADP-ribose)polymerase

Journal Article · Sun Mar 01 00:00:00 EST 1987 · Proc. Natl. Acad. Sci. U.S.A.; (United States)

DOI:https://doi.org/10.1073/pnas.84.5.1224· OSTI ID:6179292

Alkhatib, H M; Chen, D; Cherney, B; Bhatia, K; Notario, V; Giri, C; Stein, G; Slattery, E; Roeder, R G; Smulson, M E

cDNAs encoding poly(ADP-ribose) polymerase from a human hepatoma lambdagt11 cDNA library were isolated by immunological screening. One insert of 1.3 kilobases (kb) consistently hybridized on RNA gel blots to an mRNA species of 3.6-3.7 kb, which is consistent with the size of RNA necessary to code for the polymerase protein (116 kDa). This insert was subsequently used in both in vitro hybrid selection and hybrid-arrested translation studies. An mRNA species from HeLa cells of 3.6-3.7 kb was selected that was translated into a 116-kDa protein, which was selectively immunoprecipitated with anti-poly(ADP-ribose) polymerase. To confirm that the 1.3-kb insert from lambdagt11 encodes for poly(ADP-ribose) polymerase, the insert was used to screen a 3- to 4-kb subset of a transformed human fibroblast cDNA library in the Okayama-Berg vector. One of these vectors was tested in transient transfection experiments in COS cells. This cDNA insert contained the complete coding sequence for polymerase. Using pcD-p(ADPR)P as probe, it was observed that the level of poly(ADP-ribose) polymerase mRNA was elevated at 5 and 7 hr of S phase of the HeLa cell cycle, but was unaltered when artificial DNA strand breaks are introduced in HeLa cells by alkylating agents.

• Research Organization: Georgetown Univ. Schools of Medicine and Dentistry, Washington, DC
• OSTI ID: 6179292
• Journal Information: Proc. Natl. Acad. Sci. U.S.A.; (United States), Vol. 84:5
• Country of Publication: United States
• Language: English

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59 BASIC BIOLOGICAL SCIENCES
MESSENGER-RNA
HYBRIDIZATION
POLYMERASES
DNA-CLONING
RECOMBINANT DNA
CELL CYCLE
DNA REPAIR
GENE REGULATION
HELA CELLS
MAN
METHIONINE
PHOSPHORUS 32
STRAND BREAKS
SULFUR 35
AMINO ACIDS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
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DAYS LIVING RADIOISOTOPES
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DRUGS
ENZYMES
EVEN-ODD NUCLEI
ISOTOPES
LIGHT NUCLEI
LIPOTROPIC FACTORS
MAMMALS
NUCLEI
NUCLEIC ACIDS
NUCLEOTIDYLTRANSFERASES
ODD-ODD NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
PRIMATES
RADIOISOTOPES
RECOVERY
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