Rpb4 Subunit Functions Mainly in mRNA Synthesis by RNA Polymerase II

• Published in: The Journal of biological chemistry 2014-06, Vol.289 (25), p.17446-17452
• Main Authors: Schulz, Daniel, Pirkl, Nicole, Lehmann, Elisabeth, Cramer, Patrick
• Format: Article
• Language: English
• Subjects: Gene Deletion; Gene Regulation; mRNA Decay; mRNA Degradation; RNA Metabolism; RNA Polymerase II; RNA Polymerase II - genetics; RNA Polymerase II - metabolism; RNA Stability - physiology; RNA Turnover; RNA, Fungal - biosynthesis; RNA, Fungal - genetics; RNA, Messenger - biosynthesis; RNA, Messenger - genetics; Saccharomyces cerevisiae - enzymology; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae Proteins - genetics; Saccharomyces cerevisiae Proteins - metabolism; Transcription
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M114.568014

RNA polymerase II (Pol II) is the central enzyme that carries out eukaryotic mRNA transcription and consists of a 10-subunit catalytic core and a subcomplex of subunits Rpb4 and Rpb7 (Rpb4/7). Rpb4/7 has been proposed to dissociate from Pol II, enter the cytoplasm, and function there in mRNA translation and degradation. Here we provide evidence that Rpb4 mainly functions in nuclear mRNA synthesis by Pol II, as well as evidence arguing against an important cytoplasmic role in mRNA degradation. We used metabolic RNA labeling and comparative Dynamic Transcriptome Analysis to show that Rpb4 deletion in Saccharomyces cerevisiae causes a drastic defect in mRNA synthesis that is compensated by down-regulation of mRNA degradation, resulting in mRNA level buffering. Deletion of Rpb4 can be rescued by covalent fusion of Rpb4 to the Pol II core subunit Rpb2, which largely restores mRNA synthesis and degradation defects caused by Rpb4 deletion. Thus, Rpb4 is a bona fide Pol II core subunit that functions mainly in mRNA synthesis. Background: The RNA polymerase II (Pol II) subunit Rpb4 was reported to function in mRNA degradation. Results: Rpb4 deletion causes defects in mRNA synthesis and compensatory changes in mRNA degradation. Covalent attachment of Rpb4 to the Pol II core largely restores mRNA metabolism. Conclusion: Rpb4 is an integral part of Pol II and functions mainly in Pol II transcription. Significance: Rpb4 functions mainly in transcription.