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J. Biol. Chem., Vol. 279, Issue 22, 23030-23037, May 28, 2004

This Article Full Text Full Text (PDF) All Versions of this Article: 279/22/23030    most recent M400888200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Takaku, H. Articles by Ohta, A. Search for Related Content PubMed PubMed Citation Articles by Takaku, H. Articles by Ohta, A. Social Bookmarking                      What's this?

A Gcn4p Homolog Is Essential for the Induction of a Ribosomal Protein L41 Variant Responsible for Cycloheximide Resistance in the Yeast Candida maltosa^*

Hiroaki Takaku, Eishun Mutoh, Yoshiyuki Sagehashi, Ryouichi Fukuda, Hiroyuki Horiuchi, Kozo Ochi¶, Masamichi Takagi, and Akinori Ohta||

From the Department of Biotechnology, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Faculty of Applied Life Sciences, Niigata University of Pharmacy and Applied Life Sciences, Higashijima 265-1, Niitsu, Niigata 956-8603, and ^¶National Food Research Institute, 2-1-2 Kannondai, Tsukuba, Ibaraki 305-8642, Japan

Cycloheximide (CYH) resistance in the yeast Candida maltosa is based on the inducible expression of genes encoding a variant of ribosomal protein L41-Q, with glutamine at position 56 instead of the proline found in normal L41. The promoter of L41-Q2a, one of the L41-Q gene alleles encoding L41-Q, has an element similar to the Gcn4p-responsive element of Saccharomyces cerevisiae. In a previous study, this element was shown to be essential for the induction of L41-Q by CYH. In the present study, a C. maltosa GCN4 homolog, C-GCN4, was cloned. It had a long 5'-leader region with three upstream open reading frames. Enhanced expression of the C-GCN4 reporter fusion gene upon the addition of 3-aminotriazole or by mutations in start codons of all three upstream open reading frames indicates that C-GCN4 expression is under translation repression as was seen with GCN4. The C-GCN4-depleted mutant was unable to grow in a nutrient medium containing CYH and did not express L41-Q genes. Recombinant C-Gcn4p bound to the consensus DNA element for Gcn4p, 5'-(G/A)TGACTCAT-3', located upstream of L41-Q2a. Thus, C-Gcn4p, which likely functions in the general control of amino acid biosynthesis, is essential for the expression of L41-Q genes.

Received for publication, January 27, 2004 , and in revised form, March 17, 2004.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AB063247.

^* This work was performed using the facilities of the Biotechnology Research Center of The University of Tokyo. 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.

^|| To whom correspondence should be addressed. Tel.: 81-3-5841-5169; Fax: 81-3-5841-8015; E-mail: aaohta{at}mail.ecc.u-tokyo.ac.jp.

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