This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Hyman, L E Articles by Moore, C L Search for Related Content PubMed PubMed Citation Articles by Hyman, L E Articles by Moore, C L

Termination and pausing of RNA polymerase II downstream of yeast polyadenylation sites.

Department of Molecular Biology and Microbiology, Tufts University Health Sciences Campus, Boston, Massachusetts 02111-1800.

ABSTRACT

Little is known about the transcriptional events which occur downstream of polyadenylation sites. Although the polyadenylation site of a gene can be easily identified, it has been difficult to determine the site of transcription termination in vivo because of the rapid processing of pre-mRNAs. Using an in vitro approach, we have shown that sequences from the 3' ends of two different Saccharomyces cerevisiae genes, ADH2 and GAL7, direct transcription termination and/or polymerase pausing in yeast nuclear extracts. In the case of the ADH2 sequence, the RNA synthesized in vitro ends approximately 50 to 150 nucleotides downstream of the poly(A) site. This RNA is not polyadenylated and may represent the primary transcript. A similarly sized nonpolyadenylated [poly(A)-] transcript can be detected in vivo from the same transcriptional template. A GAL7 template also directs the in vitro synthesis of an RNA which extends a short distance past the poly(A) site. However, a significant amount of the GAL7 RNA is polyadenylated at or close to the in vivo poly(A) site. Mutations of GAL7 or ADH2 poly(A) signals prevent polyadenylation but do not affect the in vitro synthesis of the extended poly(A)- transcript. Since transcription of the mutant template continues through this region in vivo, it is likely that a strong RNA polymerase II pause site lies within the 3'-end sequences. Our data support the hypothesis that the coupling of this pause site to a functional polyadenylation signal results in transcription termination.

This article has been cited by other articles:

• WEST, S., ZARET, K., PROUDFOOT, N. J. (2006). Transcriptional termination sequences in the mouse serum albumin gene.. RNA 12: 655-665 [Abstract] [Full Text]  
• Ansari, A., Hampsey, M. (2005). A role for the CPF 3'-end processing machinery in RNAP II-dependent gene looping. Genes Dev. 19: 2969-2978 [Abstract] [Full Text]  
• Gillette, T. G., Gonzalez, F., Delahodde, A., Johnston, S. A., Kodadek, T. (2004). Physical and functional association of RNA polymerase II and the proteasome. Proc. Natl. Acad. Sci. USA 101: 5904-5909 [Abstract] [Full Text]  
• Nedea, E., He, X., Kim, M., Pootoolal, J., Zhong, G., Canadien, V., Hughes, T., Buratowski, S., Moore, C. L., Greenblatt, J. (2003). Organization and Function of APT, a Subcomplex of the Yeast Cleavage and Polyadenylation Factor Involved in the Formation of mRNA and Small Nucleolar RNA 3'-Ends. J. Biol. Chem. 278: 33000-33010 [Abstract] [Full Text]  
• Johanson, K., Allen, P. L., Lewis, F., Cubano, L. A., Hyman, L. E., Hammond, T. G. (2002). Saccharomyces cerevisiae gene expression changes during rotating wall vessel suspension culture. J. Appl. Physiol. 93: 2171-2180 [Abstract] [Full Text]  
• Hyman, L. E., Kwon, E., Ghosh, S., McGee, J., Chachulska, A. M. B., Jackson, T., Baricos, W. H. (2002). Binding to Elongin C Inhibits Degradation of Interacting Proteins in Yeast. J. Biol. Chem. 277: 15586-15591 [Abstract] [Full Text]  
• Graber, J. H., McAllister, G. D., Smith, T. F. (2002). Probabilistic prediction of Saccharomyces cerevisiae mRNA 3'-processing sites. Nucleic Acids Res 30: 1851-1858 [Abstract] [Full Text]  
• Gross, S., Moore, C. L. (2001). Rna15 Interaction with the A-Rich Yeast Polyadenylation Signal Is an Essential Step in mRNA 3'-End Formation. Mol. Cell. Biol. 21: 8045-8055 [Abstract] [Full Text]  
• Helden, J. v., Olmo, Marcel.l. d., Perez-Ortin, J. E. (2000). Statistical analysis of yeast genomic downstream sequences reveals putative polyadenylation signals. Nucleic Acids Res 28: 1000-1010 [Abstract] [Full Text]  
• Zhao, J., Hyman, L., Moore, C. (1999). Formation of mRNA 3' Ends in Eukaryotes: Mechanism, Regulation, and Interrelationships with Other Steps in mRNA Synthesis. Microbiol. Mol. Biol. Rev. 63: 405-445 [Abstract] [Full Text]  
• Magrath, C., Hyman, L. E. (1999). A Mutation in GRS1, a Glycyl-tRNA Synthetase, Affects 3'-End Formation in Saccharomyces cerevisiae. Genetics 152: 129-141 [Abstract] [Full Text]  
• Aranda, A., Proudfoot, N. J. (1999). Definition of Transcriptional Pause Elements in Fission Yeast. Mol. Cell. Biol. 19: 1251-1261 [Abstract] [Full Text]  
• Springer, C., Valerius, O., Strittmatter, A., Braus, G. H. (1997). The Adjacent Yeast Genes ARO4 and HIS7 Carry No Intergenic Region. J. Biol. Chem. 272: 26318-26324 [Abstract] [Full Text]  
• Kessler, M. M., Henry, M. F., Shen, E., Zhao, J., Gross, S., Silver, P. A., Moore, C. L. (1997). Hrp1, a sequence-specific RNA-binding protein that shuttles between the nucleus and the cytoplasm, is required for mRNA 3'-end formation in yeast. Genes Dev. 11: 2545-2556 [Abstract] [Full Text]