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Molecular and Cellular Biology, December 2001, p. 8203-8212, Vol. 21, No. 23
0270-7306/01/$04.00+0 DOI: 10.1128/MCB.21.23.8203-8212.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Characterization of a Six-Subunit Holo-Elongator
Complex Required for the Regulated Expression of a Group of Genes
in Saccharomyces cerevisiae
Nevan J.
Krogan and
Jack F.
Greenblatt*
Banting and Best Department of Medical
Research and Department of Molecular and Medical Genetics,
University of Toronto, Toronto, Ontario M5G 1L6, Canada
Received 9 July 2001/Accepted 22 August 2001
The Elongator complex associated with elongating RNA polymerase II
in Saccharomyces cerevisiae was originally reported to have three subunits, Elp1, Elp2, and Elp3. Using the tandem affinity purification (TAP) procedure, we have purified a six-subunit yeast Holo-Elongator complex containing three additional polypeptides, which
we have named Elp4, Elp5, and Elp6. TAP tapping and subsequent purification of any one of the six subunits result in the isolation of
all six components. Purification of Elongator in higher salt concentrations served to demonstrate that the complex could be separated into two subcomplexes: one consisted of Elp1, -2, and -3, and
the other consisted of Elp4, -5, and -6. Deletions of the individual
genes encoding the new Elongator subunits showed that only the
ELP5 gene is essential for growth. Disruption of the two
nonessential new Elongator-encoding genes, ELP4 and
ELP6, caused the same phenotypes observed with knockouts
of the original Elongator-encoding genes. Results of microarray
analyses demonstrated that the gene expression profiles of strains
containing deletions of genes encoding subunits of either Elongator
subcomplex, in which we detected significantly altered mRNA expression
levels for 96 genes, are very similar, implying that all the Elongator subunits likely function together to regulate a group of S.
cerevisiae genes in vivo.
*
Corresponding author. Mailing address: Banting and Best
Department of Medical Research, University of Toronto, 112 College St.,
Rm. 210, Toronto, Ontario, Canada M5G 1L6. Phone: (416) 978-4141. Fax:
(416) 978-8528. E-mail: jack.greenblatt{at}utoronto.ca.
Molecular and Cellular Biology, December 2001, p. 8203-8212, Vol. 21, No. 23
0270-7306/01/$04.00+0 DOI: 10.1128/MCB.21.23.8203-8212.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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