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F Sherman

Publications and source records attributed to F Sherman.

At least 163 records · Page 9Linked to original sources
PubMed1981

An extensive deletion causing overproduction of yeast iso-2-cytochrome c.

CYC7-H3 is a cis-dominant regulatory mutation that causes a 20-fold overproduction of yeast iso-2-cytochrome c. The CYC7-H3 mutation is an approximately 5 kb deletion with one breakpoint located in the 5' noncoding region of the CYC7 gene, approximately 200 base from the ATG initiation codon. The deletion apparently fuses a new regulatory region to the structural portion of the CYC7 locus. The CYC7-H3 deletion encompasses the RAD23 locus, which controls UV sensitivity and the ANP1 locus, which controls osmotic sensitivity. The gene cluster CYC7-RAD23-ANP1 displays striking similarity to the gene cluster CYC1-OSM1-RAD7, which controls, respectively, iso-1-cytochrome c, osmotic sensitivity and UV sensitivity. We suggest that these gene clusters are related by an ancient transpositional event.

Cloning, Molecular↗
PubMed1981

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

We have constructed a plasmid 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 endonuclease 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 mutational 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 leads to G change 18 nucleotides and a G leads to A change 30 nucleotides in front of the AUG initiation codon in the mRNA. The A leads to 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.

Base Sequence↗
PubMed1981

The cyc1-11 mutation in yeast reverts by recombination with a nonallelic gene: composite genes determining the iso-cytochromes c.

DNA sequence analysis of a cloned fragment directly established that the cyc1-11 mutation of iso-1-cytochrome c in the yeast Saccharomyces cerevisiae is a two-base-pair substitution that changes the CCA proline codon at amino acid position 76 to a UAA nonsense codon. Analysis of 11 revertant proteins and one cloned revertant gene showed that reversion of the cyc1-11 mutation can occur in three ways: a single base-pair substitution, which produces a serine replacement at position 76; recombination with the nonallelic CYC7 gene of iso-2-cytochrome c, which causes replacement of a segment in the cyc1-11 gene by the corresponding segment of the CYC7 gene; and either a two-base-pair substitution or recombination with the CYC7 gene, which causes the formation of the normal iso-1-cytochrome c sequence. These results demonstrate the occurrence of low frequencies of recombination between nonallelic genes having extensive but not complete homology. The formation of composite genes that share sequences from nonallelic genes may be an evolutionary mechanism for producing protein diversities and for maintaining identical sequences at different loci.

Amino Acid Sequence↗
PubMed1980

AUG is the only initiation codon in eukaryotes.

An analysis of mutants of the yeast Saccharomyces cerevisiae indicates that AUG is the sole codon capable of initiating translation of iso-1-cytochrome c. This result with yeast and the sequence results of numerous eukaryotic genes indicate that AUG is the only initiation codon in eukaryotes; in contrast, results with Escherichia coli and bacteriophages indicate that both AUG and GUG are initiation codons in prokaryotes. The difference can be explained by the lack of the t6 A hypermodified nucleoside (N-[9-(beta-D-ribofuranosyl)purin-6-ylcarbamoyl]threonine) in prokaryotic initiator tRNA and its presence in eukaryotic initiator tRNA.

Cells↗
PubMed1980

Mutants of yeast initiating translation of iso-1-cytochrome c within a region spanning 37 nucleotides.

We used a specially constructed strain, cyc1-345, of the yeast Saccharomyces cerevisiae to isolate revertants that initiated translation of iso-1-cytochrome c at various sites along an extended region of the mRNA. Normal amounts of iso-1-cytochrome c occurred when translation initiated at the abnormal sites corresponding to amino acid positions -3, -2, 3 and 5, as well as the normal position -1; 20% of the normal amounts occurred when translation initiated at the abnormal position 9. These results with cyc1-345 revertants indicate that translation of iso-1-cytochrome c can initiate with the normal efficiency at any site within the region spanning 25 nucleotides. Furthermore, because the lower amount of the short iso-1-cytochrome c in the mutant initiating at position 9 may not necessarily reflect an inefficiency of translation, we believe that translation can initiate with normal or near-normal efficiencies at any site within a 37 nucleotide region, and presumably at any site preceding and following that of the normal initiation codon. These results establish that there is no absolute requirement for a particular sequence 5' to the initiation codon, and are consistent with our previous suggestion that translation starts at the AUG codon closest to the 5' end of the mRNA.

Base Sequence↗
PubMed1980

Mating signals control expression of mutations resulting from insertion of a transposable repetitive element adjacent to diverse yeast genes.

ROAM mutations cause overproduction in S. cerevisiae. Overproduction of ROAM mutant gene products is less in MATa/MAT alpha diploid strains which cannot conjugate than in haplolid strains which can. Overproduction occurs in diploid strains capable of mating whether or not they are capable of sporulating. Overproduction decreases when haploid ROAM mutants also contain the ste7 mutation which prevents conjugation; other ste mutations do not affect the expression of ROAM mutations. Cloning of the ROAM mutant gene CYC7-H2 shows that a 5.5 kb sequence homologous to a transposable and reiterated Ty1 element is inserted in the 5' noncoding region of the CYC7 structural locus. The similar genetic properties of other ROAM mutations suggest that they each contain an inserted Ty element. These results also suggest that ROAM mutations respond to signals normally directed toward genes controlling conjugation functions, and that sequences present in Ty elements may be adjacent to structural loci and are the normal receptors for these signals.

DNA Transposable Elements↗
PubMed1980

Genes affecting the expression of cytochrome c in yeast: genetic mapping and genetic interactions.

The four mutant genes, cyc2, cyc3, cyc8 and cyc9, that affect the levels of the two iso-cytochromes c in the yeast Saccharomyces cerevisiae have been characterized and mapped. Both cyc2 and cyc3 lower the amount of iso-1-cytochrome c and iso-2-cytochrome c; whereas, cyc8 and cyc9 increase the amount of iso-2-cytochrome c. The cyc2, cyc3, cyc8 and cyc9 genes are located, respectively, on chromosomes XV, I, II and III, and are, therefore, unlinked to each other and unlinked to CYC1, the structural gene of iso-1-cytochrome c and to CYC7, the structural gene of iso-2-cytochrome c. While some cyc3 mutants are completely or almost completely deficient in cyotchromes c, none of the cyc2 mutants contained less than 10% of parental level of cytochrome c even though over one-half of the mutants contain UAA or UAG nonsense mutations. Thus, it appears as if a complete block of the cyc2 gene product still allows the formation of a residual fraction of cytochrome c. The cyc2 and cyc3 mutant genes cause deficiencies even in the presence of CYC7, cyc8 and cyc9, which normally cause overproduction of iso-2-cytochrome c. We suggest that cyc2 and cyc3 may be involved with the regulation or maturation of the iso-cytochromes c. In addition to having high levels of iso-2-cytochromes c, the cyc8 and cyc9 mutants are associated with flocculent cells and other abnormal phenotypes. The cyc9 mutant was shown to be allelic with the tup1 mutant and to share its properties, which include the ability to utilize exogenous dTMP, a characteristic flocculent morphology, the lack of sporulation of homozygous diploids and low frequency of mating and abnormally shaped cells of alpha strains. The diverse abnormalities suggest that cyc8 and cyc9 are not simple regulatory mutants controlling iso-2-cytochrome c.

Chromosome Mapping↗
PubMed1980

Dependence on mating type for the overproduction of iso-2-cytochrome c in the yeast mutant CYC7-H2.

The CYC7-H2 mutation causes an approximately 20-fold overproduction of iso-2-cytochrome c in a and alpha haploid strains of the yeast Saccharomyces cerevisiae due to an alteration in the nontranslated regulatory region that is presumably contiguous with the structural region. In this investigation, we demonstrated that heterozygosity at the mating type locus, a/alpha or a/a/alpha/alpha, prevents expression of the overproduction, while homozygosity, a/a and alpha/alpha, and hemizygosity, a/0 and alpha/0, allow full expression of the CYC7-H2 mutation, equivalent to the expression observed in a and alpha haploid strains. There is no decrease in the overproduction of iso-2-cytochrome c in a/alpha diploid strains containing either of the other two similar mutations, CYC7-H1 and CYC7-H3. It appears as if active expression of one or another of the mating-type alleles is required for the overproduction of iso-2-cytochrome c in CYC7-H2 mutants.

Base Sequence↗
PubMed1979

Primary site and second site revertants of missense mutants of the evolutionarily invariant tryptophan 64 in iso-1-cytochrome c from yeast.

The three missense mutants cyc1-132, cyc1-166 and cyc1-189 in the yeast Saccharomyces cerevisiae contain nonfunctional and thermolabile iso-1-cytochromes c and have different replacements of the tryptophan at position 64 which corresponds to the invariant tryptophan residue found in cytochromes c from all eukaryotic species. The cyc1-166 and cyc1-189 mutants contain single replacements of, respectively, serine 64 and cysteine 64, while the cyc1-132 mutant contains a double replacement of glycine 64 and alanine 65 instead of the normal tryptophan 64 and aspartic acid 65. Twenty-three intragenic revertants having at least partially functional iso-1-cytochromes c arose from these three missense mutants by single amino acid replacements of either tryptophan, phenylalanine, tyrosine or leucine at position 64, or by second-site replacements in which the mutant residues at position 64 are retained and the normal serine 45 is replaced by phenylalanine 45. Specific activities of the iso-1-cytochromes c were estimated by growth of strains on lactate medium and are as follows, in terms of the normal, for iso-1-cytochromes c altered specifically in the ways shown: 100% for phenylalanine 64; 25% for tyrosine 64; between 0 and 25% for leucine 64; 100% for phenylalanine 45, cysteine 64; 25% for phenylalanine 45, serine 64; between 0 and 25% for phenylalanine 45, glycine 64, alanine 65; and 0% for serine 64, for cysteine 64, and for glycine 64, alanine 65 iso-1-cytochromes c. The results demonstrate that small residues of glycine, serine, and cysteine at position 64 are incompatible with function; they imply that many of the 10 amino acids accessible by single base-pair substitution but not observed in primary site revertants also are incompatible with function; and they show that large hydrophobic residues of phenylalanine, leucine, and tyrosine at position 64 are capable of restoring at least partial function. The second site revertants indicate that deleterious effects of the three missense mutants can be compensated by the introduction of phenylalanine 45, which may occupy space normally filled by tryptophan 64. Altered shapes of Calpha-band spectra and at least partial instability were characteristics of all iso-1-cytochromes c found lacking tryptophan 64. Apparently, the principal role of the invariant tryptophan is stabilization of the active protein structure, by providing a large hydrophobic group at the proper location.

Amino Acid Sequence↗
PubMed1979

Mutation of the non-Mendelian suppressor, Psi, in yeast by hypertonic media.

The Psi(+) extrachromosomal determinant in the yeast Saccharomyces cerevisiae suppresses certain UAA markers and increases the efficiency of suppression of UAA suppressors and certain frameshift suppressors. Although the exact nature of Psi(+) determinant is unknown, it is believed to be a self-replicating cytoplasmic factor affecting some component of the translational machinery. In this report we describe growth conditions for efficient mutation or elimination of the Psi(+) determinant. Incubation of Psi(+) cultures in hypertonic nutrient medium resulted in rapid conversion to a culture containing predominantly Psi(-) cells during the growth cycle. The kinetics of Psi(+) to Psi(-) conversion established that the occurrence of Psi(-) cells was due to induction and not to selection of pre-existing Psi(-) cells. The results suggest that the replication of the Psi(+) determinant is sensitive to hypertonic conditions.

Journal Article↗