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PMID: 2987793 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Characterization of a factor that can prevent random transcription of cloned rDNA and its probable relationship to poly(ADP-ribose) polymerase.

Nucleic acids research ·Vol. 13 ·No. 1 ·1985-01-11 ·Pages 89-101

Kurl RN, Jacob ST

Abstract

A factor which eliminated nonspecific transcription of cloned rat rDNA was extensively purified from rat mammary adenocarcinoma ascites cells by successive fractionations on DEAE-Sephadex and heparin-Sepharose columns. The fractions containing RNA polymerase I (HS-B) and fractions eluting thereafter (HS-C) from the heparin-Sepharose column were pooled separately. Addition of HS-C to HS-B prevented random transcription of rDNA and yielded an accurate rDNA transcript with negligible non-specific transcription. The factor was essentially homogeneous and corresponded to Poly(ADP-ribose) polymerase with respect to molecular weight, dependence on DNA for its activity and its ability to undergo auto ADP-ribosylation. The total amount of protein in the transcription assay was approximately 2 micrograms, which indicates a high degree of purity of all the factors required for specific transcription of rDNA.

MeSH Terms
Adenocarcinoma/genetics Animals Cloning, Molecular DNA Topoisomerases, Type I/metabolism DNA, Ribosomal/genetics,metabolism Mammary Neoplasms, Experimental/genetics Molecular Weight NAD+ Nucleosidase/physiology Poly(ADP-ribose) Polymerases/physiology Rats Subcellular Fractions/physiology Transcription Factors/isolation & purification,pharmacology Transcription, Genetic/drug effects
Chemicals
DNA, Ribosomal Transcription Factors Poly(ADP-ribose) Polymerases NAD+ Nucleosidase DNA Topoisomerases, Type I
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Kurl R N
Jacob S T
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/13.1.89
Published
1985-01-11
Pages
89-101
Language
English
Region
England
NLM ID
0411011
PMCID
PMC340976
Subset
IM
Grants
NCI NIH HHS · CA 25078 · United States
NCI NIH HHS · CA 31894 · United States
External Links
PubMed source DOI: 10.1093/nar/13.1.89 PMC Full Text View journal details
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