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R B Wickner

Publications and source records attributed to R B Wickner.

At least 145 records · Page 8Linked to original sources
PubMed1978

Twenty-six chromosomal genes needed to maintain the killer double-stranded RNA plasmid of Saccharomyces cerevisiae.

The double-stranded RNA killer plasmid gives yeast strains carrying it both the ability to secret a protein toxin and immunity to that toxin. This report describes a new series of mutants in chromsomal genes needed for killer plasmid maintenance (mak genes). These mutants comprise 12 complementation groups. There are a total of at least 26 mak genes. Each mak gene product is needed for plasmid maintenance in diploids as well as in haploids. None of these mak mutations prevent the killer plasmid from entering the mak- spores in the process of meiotic sporulation. Complementation between mak mutants can be performed by mating meitoic spores from a makx/+ plasmid-carrying diploid with a maky haploid. If x = y, about half the diploid clones formed lose the killer plasmid. If x not equal to y, complementation occurs, and all of the diploid clones are killers.

Chromosome Mapping↗
PubMed1978

Chromosomal superkiller mutants of Saccharomyces cerevisiae.

Yeast strains carrying a 1.5 X 10(6)-dalton double-stranded RNA in virus-like particles secrete a protein toxin which is lethal to strains not carrying this species of double-stranded RNA. We find that recessive mutations in any of four chromosomal genes result in the superkiller phenotype, i.e., increased secretion of killer toxin activity by strains carrying the killer genome. These genes are designated ski1 through ski4 (for superkiller), ski3 and ski4 are located on chromosome XIV, and ski1 is on chromosome VII. A ski1 mutation results in a decreased rate of cell growth. The kex1 and kex2 mutations are epistatic to each ski mutation.

Chromosome Mapping↗
PubMed1976

Two chromosomal genes required for killing expression in killer strains of Saccharomyces cerevisiae.

The killer character of yeast is determined by a 1.4 X 10(6) molecular weight double-stranded RNA plasmid and at least 12 chromosomal genes. Wild-type strains of yeast that carry this plasmid (killers) secret a toxin which is lethal only to strains not carrying this plasmid (sensitives).--We have isolated 28 independent recessive chromosomal mutants of a killer strain that have lost the ability to secrete an active toxin but remain resistant to the effects of the toxin and continue to carry the complete cytoplasmic killer genome. These mutants define two complementation groups, kex1 and kex2. Kex1 is located on chromosome VII between ade5 and lys5. Kex2 is located on chromosome XIV, but it does not show meiotic linkage to any gene previously located on this chromosome.--When the killer plasmid of kex1 or kex2 strains is eliminated by curing with heat or cycloheximide, the strains become sensitive to killing. The mutant phenotype reappears among the meiotic segregants in a cross with a normal killer. Thus, the kex phenotype does not require an alteration of the killer plasmid.--Kex1 and kex2 strains each contain near-normal levels of the 1.4 x 10(6) molecular weight double-stranded RNA, whose presence is correlated with the presence of the killer genome.

Chromosome Mapping↗
PubMed1976

A chromosomal gene required for killer plasmid expression, mating, and spore maturation in Saccharomyces cerevisiae.

"Killer" strains of Saccharomyces cerevisiae are those that harbor a double-stranded RNA plasmid and secrete a toxin that kills only strains not carrying this plasmid (sensitives). Two chromosomal genes (kex1 and kex2) are required for the secretion of toxin by plasmid-carrying strains. The kex2 gene, which maps at a site distinct from the mating-type locus, is also required for normal mating by alpha strains and meiotic sporulation in all strains. Strains that are alpha mating-type and kex2 fail to secrete the pheromone alpha-factor or to respond to the alpha-factor II pheromone which causes a morphological change, but they do respond to alpha-factor I which causes G1 arrest in alpha cells. Strains that are alpha mating-type and kex2 show no defect in mating; pheromone secretion, or response to alpha-factor. Diploids that are homozygous for the kex2 mutation, unlike wildtype or heterozygous diploids, fail to undergo sporulation, with the defect occurring in the final spore maturation stage. These same defects in the sexual cycle are present in all kex2 mutants independent of the presence of the "killer" plasmid.

Animals↗
PubMed1976

Mutants of the killer plasmid of Saccharomyces cerevisiae dependent on chromosomal diploidy for expression and maintenance.

Mutants of the killer plasmid of Saccharomyces cerevisiae have been isolated that depend upon chromosomal diploidy for the expression of plasmid functions and for replication or maintenance of the plasmid itself. These mutants are not defective in any chromosomal gene needed for expression or replication of the killer plasmid.--Haploids carrying these mutant plasmids (called d for diploid-depen;ent) are either unable to kill or unable to resist being killed or both and show frequent loss of the plasmid. The wild-type phenotype (K+R+) is restored by mating the d plasmid-carrying strain with either (a) a wild-type sensitive strain which apparently has no killer plasmid; (b) a strain which has been cured of the killer plasmid by growth at elevated temperature; (c) a strain which has been cured of the plasmid by growth in the presence of cycloheximide; (d) a strain which has lost the plasmid because it carries a mutation in a chromosomal mak gene; or (e) a strain of the opposite mating type which carries the same d plasmid and has the same defective phenotype, indicating that the restoration of the normal phenotype is not due to recombination between plasmid genomes or complementation of plasmid or chromosomal genes.--Sporulation of the phenotypically K+R+ diploids formed in matings between d and wild-type nonkiller strains yields tetrads, all four of whose haploid spores are defective for killing or resistance or maintenance of the plasmid or a combination of these. Every defective phenotype may be found among the segregants of a single diploid clone carrying a d plasmid. These defective segregants resume the normal killer phenotype in the diploids formed when a second round of mating is performed, and the segregants from a second round of meiosis and sporulation are again defective.

Crosses, Genetic↗
PubMed1974

Chromosomal and nonchromosomal mutations affecting the "killer character" of Saccharomyces cerevisiae.

The "killer character" of Saccharomyces cerevisiae is a nonchromosomal genetic element which imparts to cells carrying it (a) the ability to kill cells which lack it, by secreting the soluble macromolecular killer substance and (b) the ability to resist the killing effects of the killer substance.-Mutants have been isolated from S. cerevisiae carrying the "killer character". Mutants were found in two nuclear genes (mak1 and mak2) involved in maintenance of the genetic element of the killer character. Mutants in three other nuclear genes could maintain the cytoplasmic genetic element normally; two of these were able to express resistance to killing, but not the ability to kill (kex1 and kex2), while the third could express the killing function, but was sensitive to killing and is thus a "suicide" strain (rex1). These five nuclear genes were all distinct.-Several mutants were detected which showed non-mendelian segregation indicating mutation of the "cytoplasmic" genetic element. Some such mutants had normal resistance to killing but were either unable to kill at all or showed marked reduction in the ability to kill. Others had lost both killing ability and resistance. None were suppressive.-Two nonkilling mutants, both of which showed cytoplasmic inheritance, were mated. A large, but variable, proportion of diploid colonies, plated after several generations of growth, showed normal killing which has then stably inherited on further subcloning.

Cell Nucleus↗
PubMed1974

Mutants of Saccharomyces cerevisiae that incorporate deoxythymidine-5'-monophosphate into deoxyribonucleic acid in vivo.

Spontaneous mutants of Saccharomyces cerevisiae able to incorporate deoxythymidine-5'-monophosphate (dTMP) into deoxyribonucleic acid (DNA) have been selected based on their ability to grow in the presence of aminopterin and sulfanilamide if dTMP is present. Essentially all mutants (called tup) selected in this way required dTMP for growth in the presence of the two drugs, but none required dTMP in the absence of the drugs. Neither thymine nor thymidine would satisfy this requirement. Equimolar amounts of (32)P- and (3)H-base-labeled dTMP were incorporated by the mutants into alkali-stable, deoxyribonuclease-sensitive material. In the presence of aminopterin and sulfanilamide, this incorporation was sufficient to account for a substantial proportion of the thymine residues in the cellular DNA, whereas in the absence of the drugs only about 40% as much of the thymine residues originated from the medium. Of 29 mutants examined, all were recessive and 17 showed 2:2 segregation in crosses with a wild-type strain. The lesions in these mutants fell into four complementation groups: one (tup1) occurs on chromosome III; another (tup3) is on chromosome II; and a third (tup4) was centromere linked. Strains of the genotype alpha tup1 mated with lower than normal efficiency with a strains, but with higher than normal efficiency with alpha strains. Strains of genotype a/alpha tup1/tup1 failed to sporulate, whereas homozygous diploids for tup2, tup3, or tup4 sporulated normally, as did a/alpha tup1/+ strains.

Aminopterin↗