Journal
JOURNAL OF MOLECULAR BIOLOGY
Volume 404, Issue 1, Pages 1-15Publisher
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/j.jmb.2010.09.040
Keywords
Iws1; RNAPII; transcription; preloaded promoter
Categories
Funding
- National Institutes of Health [GM067777]
- National Science Foundation [MCB-0843073]
- Howard Hughes Medical Institute
- Div Of Molecular and Cellular Bioscience
- Direct For Biological Sciences [0843073] Funding Source: National Science Foundation
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Spn1/Iws1 plays essential roles in the regulation of gene expression by RNA polymerase II (RNAPII), and it is highly conserved in organisms ranging from yeast to humans. Spn1 physically and/or genetically interacts with RNAPII, TBP (TATA-binding protein), TFIIS (transcription factor ITS), and a number of chromatin remodeling factors (Swi/Snf and Spt6). The central domain of Spn1 (residues 141-305 out of 410) is necessary and sufficient for performing the essential functions of SPN1 in yeast cells. Here, we report the high-resolution (1.85 angstrom) crystal structure of the conserved central domain of Saccharomyces cerevisiae Spn1. The central domain is composed of eight alpha-helices in a right-handed superhelical arrangement and exhibits structural similarity to domain I of THIS. A unique structural feature of Spn1 is a highly conserved loop, which defines one side of a pronounced cavity. The loop and the other residues forming the cavity are highly conserved at the amino acid level among all Spn1 family members, suggesting that this is a signature motif for Spn1 orthologs. The locations and the molecular characterization of temperature-sensitive mutations in Spn1 indicate that the cavity is a key attribute of Spn1 that is critical for its regulatory functions during RNAPII-mediated transcriptional activity. (C) 2010 Elsevier Ltd. All rights reserved.
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