Biosynthesis of the human DNA repair enzyme uracil DNA glycosylase

Conference · Fri May 01 00:00:00 EDT 1987 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:6023360

Vollberg, T M; Cool, B L; Sirover, M A

Anti-human uracil DNA glycosylase monoclonal antibodies have been used to study the in vitro and in vivo biosynthesis of this human base excision repair enzyme. The in vitro translation of poly(A+)uracil DNA glycosylase mRNA was examined by immunoprecipitation of the (/sup 35/S)methionine-labeled translation products. As identified by sucrose density analysis, immunoreactive in vitro products of 37,000 daltons and 24,000 daltons were translated from 16S poly(A+)RNA and from 11S poly (A+)RNA, respectively. However, only the 37,000 Mr polypeptide could be detected by immunoblot analysis of crude human placental and fibroblast extracts or by immunoprecipitation of (/sup 35/S) labeled normal human cell extracts. This relative molecular weight correspond to the molecular weight observed for homogeneous human placental uracil DNA glycosylase. A placental mRNA preparation was used to construct a cDNA library in pUC8. Immunological screening identified two recombinant DNA plasmids each containing a 340 base pair insert. Northern blot analysis demonstrated that the insert hybridized to a 16S poly (A+)mRNA. These results suggest that immunoreactive uracil DNA glycosylase protein was synthesized at its enzymatically active molecular weight as the primary translation product of a 16S poly (A+)mRNA.

• Research Organization: Temple Univ. School of Medicine, Philadelphia, PA
• OSTI ID: 6023360
• Report Number(s): CONF-870644-
• Journal Information: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Vol. 46:6; ISSN FEPRA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
ANTIBODIES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMISTRY
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
BIOSYNTHESIS
CELL CONSTITUENTS
CHEMISTRY
DAYS LIVING RADIOISOTOPES
DNA
DNA REPAIR
ENZYMES
EVEN-ODD NUCLEI
GLYCOSYL HYDROLASES
HYDROLASES
IMMUNOASSAY
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
MAMMALS
MAN
MESSENGER-RNA
MOLECULAR WEIGHT
MONOCLONAL ANTIBODIES
NUCLEI
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PLASMIDS
PRIMATES
RADIOISOTOPES
RECOMBINANT DNA
RECOVERY
REPAIR
RNA
SULFUR 35
SULFUR ISOTOPES
SYNTHESIS
TRACER TECHNIQUES
VERTEBRATES