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PMID: 3025649 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Structure of the Saccharomyces cerevisiae HO gene and analysis of its upstream regulatory region.

Molecular and cellular biology ·Vol. 6 ·No. 12 ·1986-12-00 ·Pages 4281-94

Russell DW, Jensen R, Zoller MJ, Burke J, Errede B, Smith M, Herskowitz I

Abstract

The HO gene product of Saccharomyces cerevisiae is a site-specific endonuclease that initiates mating type interconversion. We have determined the nucleotide sequence of a 3,129-base-pair (bp) segment containing HO. The segment contains a single long open reading frame encoding a polypeptide of 586 amino acids, which has unusual (unbiased) codon usage and is preceded by 762 bp of upstream region. The predicted HO protein is basic (16% lysine and arginine) and is calculated to have a secondary structure that is 30% helical. The corresponding transcript is initiated approximately 50 nucleotides prior to the presumed initiation codon. Insertion of an Escherichia coli lacZ gene fragment into the putative HO coding segment inactivated HO and formed a hybrid HO-lacZ gene whose beta-galactosidase activity was regulated by the mating type locus in the same manner as HO (repressed by a 1-alpha 2). Upstream regions of 1,360 and 762 bp conferred strong repression; 436 bp led to partial constitutivity and 301 bp to full constitutivity. Thus, DNA sequences that confer repression of HO by a1-alpha 2 are at least 250 nucleotides upstream of the transcription start point and are within 436 nucleotides of the HO initiation codon. The progressive loss of repression suggests that both the -762 to -436 and the -436 to -301 intervals contain sites for regulation by a1-alpha 2. The HO gene contains two distinct regions that promote autonomous replication of plasmids in S. cerevisiae. These regions contain sequences that are homologous to the two conserved sequences that are associated with ARS activity.

MeSH Terms
Amino Acid Sequence Base Sequence DNA Restriction Enzymes/genetics Deoxyribonucleases, Type II Site-Specific Genes Genes, Fungal Genes, Mating Type, Fungal Genes, Regulator Genotype Saccharomyces cerevisiae/enzymology,genetics Saccharomyces cerevisiae Proteins beta-Galactosidase/genetics
Chemicals
Saccharomyces cerevisiae Proteins DNA Restriction Enzymes HO protein, S cerevisiae SCEI protein, S cerevisiae Deoxyribonucleases, Type II Site-Specific beta-Galactosidase
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Russell D W
Jensen R
Zoller M J
Burke J
Errede B
Smith M
Herskowitz I
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.6.12.4281-4294.1986
Published
1986-12-00
Pages
4281-94
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC367210
Subset
IM
Databases
GENBANK
M14678
External Links
PubMed source DOI: 10.1128/mcb.6.12.4281-4294.1986 PMC Full Text View journal details
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