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Nucleic Acids Research, 1985, Vol. 13, No. 1 89-101
© 1985

Articles

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

Rabinder N. Kurl and Samson T. Jacob

Department of Pharmacology, Milton S. Hershey Medical Center, Pennsylvania State University, College of Medicine Hershey, PA 17033, USA

Received August 27, 1984. Revised November 30, 1984. Accepted November 30, 1984.

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 (H3-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 homogenous 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 µg, which indicates a high degree of purity of all the factors required for specific transcription of rDNA.


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