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The C‐terminal domain of the Spt16 subunit of FACT affects transcription elongation
Author(s) -
Stevens Jennifer R.,
O’Donnell Allyson F.,
Johnston Gerald C.,
Singer Richard A.
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.4.a468-b
Subject(s) - elongation , protein subunit , terminal (telecommunication) , domain (mathematical analysis) , microbiology and biotechnology , transcription (linguistics) , chemistry , biology , gene , computer science , biochemistry , computer network , mathematics , philosophy , mathematical analysis , materials science , linguistics , ultimate tensile strength , metallurgy
In the budding yeast S. cerevisiae , the abundant nuclear complex FACT ( F acilitates C hromatin T ransactions) is a heterodimer of two essential proteins: Spt16 and Pob3. FACT is highly conserved among eukaryotes, and has been implicated in transcription, DNA replication and DNA repair. To learn more about the roles FACT plays in these processes, genome‐wide synthetic‐lethal analysis of several mutant alleles of SPT16 was carried out. These mutant alleles were all identified by a dominant Spt ¯ phenotype, yet they yielded radically different patterns of genetic interactions. Those alleles that display interactions with genes involved in transcription elongation all contain mutations affecting the C‐terminal portion of Spt16; this clustering of mutations may identify a region of Spt16 responsible for the proposed histone‐chaperone activity of FACT. Further studies with one mutant allele that affects the C‐terminal domain, spt16‐E857K , demonstrate that this mutant protein interacts more weakly with histone H2B than does wild‐type Spt16 or a mutant Spt16 without C‐terminal‐domain alterations. We therefore propose a model in which weakened interactions between FACT and histones negatively affects transcription elongation. This work has been supported by the Natural Sciences and Engineering Research Council (NSERC) and the Canadian Institutes for Health Reseach (CIHR).