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

Rabinder N. Kurl | Zendy; Samson T. Jacob | Zendy

Open Access

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

Author(s) -

Rabinder N. Kurl,

Samson T. Jacob

Publication year - 1985

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/13.1.89

Subject(s) - biology , transcription (linguistics) , microbiology and biotechnology , polymerase , rna polymerase , dna , transcription factor , rna polymerase ii , sepharose , rna polymerase i , promoter , rna , biochemistry , gene , gene expression , enzyme , rna dependent rna polymerase , philosophy , linguistics

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.

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