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J. Biol. Chem., Vol. 278, Issue 40, 39037-39043, October 3, 2003
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Molecular Evidence That the Eukaryotic THO/TREX Complex Is Required for Efficient Transcription Elongation*
From the Departamento de Genética, Facultad de Biología, Universidad de Sevilla, Avenida Reina Mercedes 6, 41012 Sevilla, Spain
THO/TREX is a conserved eukaryotic complex formed by the core THO complex plus proteins involved in mRNA metabolism and export such as Sub2 and Yra1. Mutations in any of the THO/TREX structural genes cause pleiotropic phenotypes such as transcription impairment, increased transcription-associated recombination, and mRNA export defects. To assay the relevance of THO/TREX complex in transcription, we performed in vitro transcription elongation assays in mutant cell extracts using supercoiled DNA templates containing two G-less cassettes. With these assays, we demonstrate that hpr1
, tho2, and mft1 mutants of the THO complex and sub2 mutants show significant reductions in the efficiency of transcription elongation. The mRNA expression defect of hpr1 mutants was not due to an increase in mRNA decay, as determined by mRNA half-life measurements and mRNA time course accumulation experiments in the absence of Rrp6p exoribonuclease. This work demonstrates that THO and Sub2 are required for efficient transcription elongation, providing further evidence for the coupling between transcription and mRNA metabolism and export.
Received for publication, June 2, 2003 , and in revised form, July 7, 2003.
* This work was supported by Spanish Ministry of Science and Technology Grant BMC2000-0439 and the Human Frontier Science Program Grant RG1999/0075. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
Recipient of a predoctoral training grant from the Spanish Ministry of Education.
Recipient of a predoctoral training grant from the Spanish Ministry of Health.
¶ To whom correspondence should be addressed. Tel.: 34-954-557107; Fax: 34-954-557104; E-mail: aguilo{at}us.es.
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