DNA sequence of a mutation in the leader region of the yeast iso-1-cytochrome c mRNA

Journal Article · Wed Jul 01 04:00:00 EDT 1981 · Cell; (United States)

DOI:https://doi.org/10.1016/0092-8674(81)90253-1· OSTI ID:6844518

Stiles, J I ^[1]; Szostak, J W; Young, A T; Wu, R; Consaul, S; Sherman, F

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1. Indiana State Univ., Terre Haute

A plasmid was constructed that selectively integrates adjacent to the CYC1 locus, which determines iso-1-cytochrome c in the yeast Saccharomyces cerevisiae. Different CYC1 alleles can be conveniently recovered by digestion of total DNA from transformed strains with BgI II, a restriction endouclease that does not cut the vector or the CYC1 gene, followed by transformation of Escherichia coli, selecting the ampicillin resistance gene carried on the original vector. This procedure was used to clone the cyc1-362 gene, which contains an alteration in front of the AUG initiation codon. The cyc1-362 mutation causes a deficiency of the iso-1-cytochrome c protein but still allows transcription of the iso-1-cytochrome c mRNA. DNA sequence analysis showed that the cyc1-362 mutation consisted of two single-base-pair substitutions, producing an A ..-->.. G change 18 nucleotides and a G ..-->.. A change 30 nucleotides in front of the AUG initiation codon in the mRNA. The A ..-->.. G change at position -18 resulted in the creation of an AUG triplet, which is proximal to the normal initiation site and out of phase with the normal reading frame. The deficiency of iso-1-cytochrome c is most simply explained by assuming that translation initiates at the more proximal abnormal AUG site but not at the normal AUG site.

• OSTI ID: 6844518
• Journal Information: Cell; (United States), Journal Name: Cell; (United States) Vol. 25; ISSN CELLB
• Country of Publication: United States
• Language: English

Related Subjects

550200 -- Biochemistry
550400* -- Genetics
59 BASIC BIOLOGICAL SCIENCES
BACTERIA
CELL CONSTITUENTS
CLONING
CYTOCHROMES
DNA
DNA REPLICATION
DNA-CLONING
ENZYMES
ESCHERICHIA COLI
FUNGI
GENES
GENETIC MAPPING
MAPPING
MICROORGANISMS
MUTATIONS
NUCLEIC ACID REPLICATION
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PIGMENTS
PLANTS
PLASMIDS
RNA
SACCHAROMYCES
SACCHAROMYCES CEREVISIAE
YEASTS