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Molecular and Cellular Biology, February 2007, p. 937-948, Vol. 27, No. 3
0270-7306/07/$08.00+0     doi:10.1128/MCB.01584-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Nucleolin Is Required for RNA Polymerase I Transcription In Vivo{triangledown}

Brenden Rickards,1 S. J. Flint,1 Michael D. Cole,2 and Gary LeRoy1*

Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544,1 Department of Pharmacology and Toxicology and Department of Genetics, Dartmouth Medical School, Lebanon, New Hampshire 037562

Received 24 August 2006/ Returned for modification 4 October 2006/ Accepted 14 November 2006

Eukaryotic genomes are packaged with histones and accessory proteins in the form of chromatin. RNA polymerases and their accessory proteins are sufficient for transcription of naked DNA, but not of chromatin, templates in vitro. In this study, we purified and identified nucleolin as a protein that allows RNA polymerase II to transcribe nucleosomal templates in vitro. As immunofluorescence confirmed that nucleolin localizes primarily to nucleoli with RNA polymerase I, we demonstrated that nucleolin allows RNA polymerase I transcription of chromatin templates in vitro. The results of chromatin immunoprecipitation experiments established that nucleolin is associated with chromatin containing rRNA genes transcribed by RNA polymerase I but not with genes transcribed by RNA polymerase II or III. Knockdown of nucleolin by RNA interference resulted in specific inhibition of RNA polymerase I transcription. We therefore propose that an important function of nucleolin is to permit RNA polymerase I to transcribe nucleolar chromatin.

* Corresponding author. Mailing address: Department of Molecular Biology, Princeton University, Princeton, NJ 08544. Phone: (609) 258-5414. Fax: (609) 258-4575. E-mail: gleroy{at}princeton.edu.

{triangledown} Published ahead of print on 27 November 2006.

Molecular and Cellular Biology, February 2007, p. 937-948, Vol. 27, No. 3
0270-7306/07/$08.00+0     doi:10.1128/MCB.01584-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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