A MAPPING METHOD FOR SACCHAROMYCES CEREVISIAE USING rad52-INDUCED CHROMOSOME LOSS

Genetics ◽  
1985 ◽  
Vol 110 (4) ◽  
pp. 569-589
Author(s):  
David Schild ◽  
Robert K Mortimer
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Mapping Method ◽  
Treatment Result ◽  
Recessive Mutation ◽  
Chromosome Loss ◽  
Chromosome 2 ◽  
Chromosome X ◽  
Coordinate Expression ◽  
Chromosomal Markers ◽  
The Right

ABSTRACT Saccharomyces cerevisiae diploids homozygous for the rad52-1 mutation have previously been shown to lose chromosomes mitotically. Spontaneous events and events following low levels of X-ray or methyl methanesulfonate treatment result in monosomic diploids, whereas higher levels of treatment result in near haploidization. This rad52-1-dependent chromosome loss has been used to develop a new mapping method which can be used to assign a previously unmapped gene to a chromosome. Chromosome loss mapping can be done in either of two ways: (1) if a diploid, homozygous for rad52-1 but heterozygous for a variety of other recessive markers, is constructed with an unmapped recessive mutation in coupling with known chromosomal markers, chromosome loss will result in the coordinate expression of the mutation and other recessive markers on the same chromosome; (2) if, however, the diploid is constructed with the unmapped mutation in repulsion to chromosomal markers, then even haploidization will never result in the coordinate expression of the unmapped mutation and other markers on the same homologous chromosome pair—This mapping method and subsequent tetrad analyses have been used to locate hom6 on chromosome X, ade4 on chromosome XIII and cdc31 on chromosome XV and to demonstrate that met5, previously assigned to chromosome V, actually maps to chromosome X; the met- marker on chromosome V has been shown to be met6. GAL80 and SUP5, previously assigned to an unmapped fragment, have now been mapped to the right arm of chromosome XIII.

Mitotic chromosome loss induced by methyl benzimidazole-2-yl-carbamate as a rapid mapping method in Saccharomyces cerevisiae

1982 ◽  
Vol 2 (9) ◽  
pp. 1080-1087
Author(s):  
J S Wood
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Mitotic Chromosome ◽  
Chromosomal Location ◽  
Mapping Method ◽  
Chromosome Loss ◽  
Diploid Strain ◽  
Simple Method ◽  
Rapid Mapping

Mitotic chromosome loss induced by methyl benzimidazole-2-yl-carbamate has been utilized as a rapid and simple method for assigning genes to individual chromosomes in Saccharomyces cerevisiae. This technique relied on the segregation of heterozygous markers in a diploid strain after methyl benzimidazole-2-yl-carbamate treatment due to loss of whole chromosomes. Correlations between the expression of an unmapped gene and that of a previously mapped recessive marker indicated chromosomal linkage. Depending on whether the unmapped gene and the marker were located in coupling or in repulsion, either positive or negative correlations were seen. The chromosomal location of several previously mapped genes were confirmed as a test of the method, and one previously unmapped gene, nib1, was mapped.

scholarly journals Isolation of the gene encoding the Saccharomyces cerevisiae centromere-binding protein CP1.

1990 ◽  
Vol 10 (6) ◽  
pp. 2458-2467 ◽  
Author(s):  
R E Baker ◽  
D C Masison
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Gene Disruption ◽  
Mitotic Chromosome ◽  
Binding Protein ◽  
Chromosome Loss ◽  
Chromosome X ◽  
Gene Encoding ◽  
Yeast Saccharomyces Cerevisiae ◽  
Unexpected Consequence ◽  
Cell Doubling Time

CP1 is a sequence-specific DNA-binding protein of the yeast Saccharomyces cerevisiae which recognizes the highly conserved DNA element I (CDEI) of yeast centromeres. We cloned and sequenced the gene encoding CP1. The gene codes for a protein of molecular weight 39,400. When expressed in Escherichia coli, the CP1 gene directed the synthesis of a CDEI-binding protein having the same gel mobility as purified yeast CP1. We have given the CP1 gene the genetic designation CEP1 (centromere protein 1). CEP1 was mapped and found to reside on chromosome X, 2.0 centimorgans from SUP4. Strains were constructed in which most of CEP1 was deleted. Such strains lacked detectable CP1 activity and were viable; however, CEP1 gene disruption resulted in a 35% increase in cell doubling time and a ninefold increase in the rate of mitotic chromosome loss. An unexpected consequence of CP1 gene disruption was methionine auxotrophy genetically linked to cep1. This result and the recent finding that CDEI sites in the MET25 promoter are required to activate transcription (D. Thomas, H. Cherest, and Y. Surdin-Kerjan, J. Mol. Biol. 9:3292-3298, 1989) suggest that CP1 is both a kinetochore protein and a transcription factor.

scholarly journals Mitotic chromosome loss induced by methyl benzimidazole-2-yl-carbamate as a rapid mapping method in Saccharomyces cerevisiae.

1982 ◽  
Vol 2 (9) ◽  
pp. 1080-1087 ◽  
Author(s):  
J S Wood
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Mitotic Chromosome ◽  
Chromosomal Location ◽  
Mapping Method ◽  
Chromosome Loss ◽  
Diploid Strain ◽  
Simple Method ◽  
Rapid Mapping

Mitotic chromosome loss induced by methyl benzimidazole-2-yl-carbamate has been utilized as a rapid and simple method for assigning genes to individual chromosomes in Saccharomyces cerevisiae. This technique relied on the segregation of heterozygous markers in a diploid strain after methyl benzimidazole-2-yl-carbamate treatment due to loss of whole chromosomes. Correlations between the expression of an unmapped gene and that of a previously mapped recessive marker indicated chromosomal linkage. Depending on whether the unmapped gene and the marker were located in coupling or in repulsion, either positive or negative correlations were seen. The chromosomal location of several previously mapped genes were confirmed as a test of the method, and one previously unmapped gene, nib1, was mapped.

scholarly journals MAPPING AND GENE CONVERSION STUDIES WITH THE STRUCTURAL GENE FOR ISO-1-CYTOCHROME C IN YEAST

Genetics ◽  
1975 ◽  
Vol 81 (4) ◽  
pp. 615-629
Author(s):  
Christopher W Lawrence ◽  
Fred Sherman ◽  
Mary Jackson ◽  
Richard A Gilmore
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Gene Conversion ◽  
The Other ◽  
Crossing Over ◽  
Wild Type ◽  
Chromosome X ◽  
Tetrad Analysis ◽  
Yeast Saccharomyces Cerevisiae ◽  
Distal Marker ◽  
The Right

ABSTRACT We have investigated the order of the four genes cyc1, rad7, SUP4, and cdc8 which form a tightly linked cluster on the right arm of chromosome X in the yeast Saccharomyces cerevisiae. Crossing over and coconversion data from tetrad analysis established the gene order to be centromere–cyc1–rad7–SUP4. Also cdc8 appeared to be distal to SUP4 on the basis of crossovers that were associated with conversion of SUP4. The frequencies of recombination and the occurrence of coconversions suggest that these four genes are contiguous or at least nearly so. Gene-conversion frequencies for several cyc1 alleles were studied, including cyc1–1, a deletion of the whole gene that extends into the rad7 locus. The cyc1–1 deletion was found to be capable of conversion, though at a frequency some fivefold less than the other alleles studied, and both 3:1 and 1:3 events were detected. In general 1:3 and 3:1 conversion events were equally frequent at all loci studied, and approximately 50% of conversions were accompanied by reciprocal recombination for flanking markers. The orientation of the cyc1 gene could not be clearly deduced from the behavior of the distal marker SUP4 in wild-type recombinants that arose from diploids heteroallelic for cyc1 mutations.

Chromosomal assignment of mutations by specific chromosome loss in the yeast Saccharomyces cerevisiae.

Genetics ◽  
1990 ◽  
Vol 125 (2) ◽  
pp. 333-340 ◽  
Author(s):  
L P Wakem ◽  
F Sherman
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Plasmid Dna ◽  
High Frequency ◽  
Recessive Mutation ◽  
Chromosomal Assignment ◽  
Chromosome Loss ◽  
Specific Chromosome ◽  
Yeast Saccharomyces Cerevisiae ◽  
Homologous Chromosomes

Abstract Yeast 2-microns plasmids were integrated near the centromere of a different chromosome in each of 16 cir0 mapping strains of Saccharomyces cerevisiae. The specific chromosomes containing the integrated 2-microns plasmid DNA were lost at a high frequency after crossing the cir0 strains to cir+ strains. A recessive mutation in a cir+ strain can then be easily assigned to its chromosome using this set of mapping strains, since the phenotype of the recessive mutation will be manifested only in diploids having the integrated 2-microns plasmid and the unmapped mutation on homologous chromosomes.

scholarly journals Reciprocal Chromosome Translocation Between the Left-End 220kb of Chromosome II and the Right-End 270kb of Chromosome X in Saccharomyces cerevisiae

2018 ◽  
Vol 10 (4) ◽  
pp. 1
Author(s):  
Masaharu Takeda ◽  
Takahito Okushiba
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Chromosome Translocation ◽  
Original Strain ◽  
Chromosome X ◽  
A Cell ◽  
Colony Color ◽  
The Stability ◽  
The Right ◽  
Chromosome Ii ◽  
Reciprocal Chromosome Translocation

Southern hybridization of chromosomes and the physical mapping of the genes used as several probes on the respective chromosomes II and X showed that the left-end ca. 220kb of chromosome II including ATP1 was exchanged the right-end ca. 270kb of chromosome X including ATP2 resulting the reciprocal chromosome translocation in the yeast strain YNN290, Saccharomyces cerevisiae. YTO290, the mutated strain by the reciprocal chromosome translocation as above described, was changed from red to white of the colony-color, and sizes of chromosome II lengthened from ca. 830kb to ca. 900kb and chromosome X shortened from ca. 760kb to ca. 690kb, respectively, in compared with the original strain YNN290. But, YTO290 strain was the same as the original strain YNN290 for other properties; the nutrient requiring of the genotype, the ploidy, the mitochondrial respiratory activity, the cell-size, and the growth-rate (doubling time), the number of chromosomes in a cell, It should be as a total number of nucleotides (bases) of genome.ATP1 or ATP2 and their neighboring base sequences respectively should be transferred from chromosome II left-end ca. 220kb to chromosome X right-end or chromosome X right-end ca. 270kb to chromosome II left-end accompanying with this reciprocal chromosome translocation. This mutated (the reciprocal chromosomes II and X translocation = exchanged those end-sequences as above described) strain, YTO290, seemed to lead to decrease the stability of the changed chromosomes II and X. The mutated strain, YTO290 might be observed to go back to the respective chromosomes II and X of the original strain, YNN290, in several months later even at 4°C.

scholarly journals Chromosomes XIV and XVII of Saccharomyces cerevisiae constitute a single linkage group.

1982 ◽  
Vol 2 (11) ◽  
pp. 1399-1409 ◽  
Author(s):  
S Klapholz ◽  
R E Esposito
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Linkage Group ◽  
Mapping Method ◽  
Normal Chromosome ◽  
Tetrad Analysis ◽  
Single Linkage ◽  
Previous Evidence ◽  
The Right ◽  
Lines Of Evidence

We present several lines of evidence that chromosomes XIV and XVII of Saccharomyces cerevisiae are not independent chromosomes, but rather constitute a single linkage group. Studies which made use of a new mapping method based on the haploidization-without-recombination meiotic phenotype of the spoll mutant initially indicated that markers on chromosomes XIV and XVII were linked. Tetrad analysis was used to establish gene-gene distances, and a new chromosome XIV map incorporating markers originally assigned to chromosome XVII was derived. During the course of trisomic segregation studies, we discovered that a 2n + 2 homothallic diploid, originally believed to be tetrasomic for chromosome XVII (now XIV), carries two normal chromosome XIV homologs and two aberrant homologs which appear to be deficient for a large portion of the right arm of XIV. The previous evidence that established chromosome XVII as an independent linkage group is discussed in the light of these findings.

scholarly journals Segregation of centric Y-autosome translocations in Drosophila melanogaster: I. Segregation determinants in males

1985 ◽  
Vol 45 (1) ◽  
pp. 51-79 ◽  
Author(s):  
Raphael Falk ◽  
Shula Baker ◽  
Ana Rahat
Keyword(s):  
Drosophila Melanogaster ◽  
Sex Chromosome ◽  
Chromosome 2 ◽  
Chromosome X ◽  
Complete Independence ◽  
Group 2 ◽  
The Right ◽  
Complete Dependence ◽  
Group 1 ◽  
Chromosome Disjunction

SummaryA special screening procedure for the detection of induced Y-autosome translocations with centric breakpoints was applied. A series of Experimental stocks was constructed, each containing a different half of one of the induced T(Y; 2)'s (T element). The three other elements that were involved in the segregation experiments in each stock were a sex chromosome (X element), an inverted chromosome 2 (A element), and a free arm of chromosome 2 (F element). It is not feasible to determine the relative frequencies of all the 16 possible gamete types by mating an Experimental stock to one tester, nor to different testers that have each at least one class of progeny of the same genotype. Each Experimental stock was mated to four different Tester stocks and the data were calibrated so that a coherent segregation pattern could be obtained.Segregation patterns in meiosis of males from 15 Experimental stocks, each with a different T element were studied. In most Experimental stocks the T element was of the left autosomal arm, while the F element was of the right autosomal arm. In four Experimental stocks the X element segregated independently of the A, F and T elements. In these Group 1 stocks, both the F and the T elements disjoined regularly from the A element. It was concluded that the T element of these stocks had no sex-chromosome disjunction determinants (‘S-determinants’) to interact with the determinants on the X element. Both the T elements and the F elements carried autosomal disjunction determinants (‘H-determinants’) that secured the segregation of the autosomal elements. The H-determinants of the left autosomal arm were qualitatively different from those of the right arm.In the remaining 11 Group-2 Experimental stocks the X and T elements disjoined regularly, indicating the presence of S-determinants on the T elements of these stocks. The segregation of the T and the A elements in these stocks varied from nearly complete dependence to complete independence. It was concluded that this gradation reflected differences in the quantity of H-determinants present on the T elements of these Experimental stocks. It was impossible to discriminate between a model of continuous H determinants activity and one of a finite discrete number of determinants. The results do not agree with the claim that there are no autosomal disjunction determinants in the proximal heterochromatin of chromosome 2.The S-determinants on the BsYy+ chromosome were located both adjacent to the centromere and distally on the long arm. The latter were probably translocated to the Y chromosome together with the Bs marker.

A NEW MAPPING METHOD EMPLOYING A MEIOTIC REC- MUTANT OF YEAST

Genetics ◽  
1982 ◽  
Vol 100 (3) ◽  
pp. 387-412 ◽  
Author(s):  
Sue Klapholz ◽  
Rochelle Easton Esposito
Keyword(s):  
Drug Resistance ◽  
Chromosome Segregation ◽  
Mapping Method ◽  
Recessive Mutation ◽  
Drug Resistant ◽  
Resistance Marker ◽  
Spore Viability ◽  
Mapping Procedure ◽  
Chromosomal Markers ◽  
Drug Resistance Marker

ABSTRACT A rapid new mapping method has been developed for localizing a dominant or recessive mutation to a particular chromosome of yeast. The procedure utilizes the ability of strains homozygous for the spo11-1 mutation to undergo chromosome segregation without appreciable recombination during sporulation. The level of sporulation in spo11-1/spo11-1 diploids is reduced and asci are often immature or abnormal in appearance; spore viability is less than 1%. The first step of the mapping procedure is the construction of a haploid spo11-1 strain carrying a recessive drug-resistance marker and the unmapped mutation(s). This strain is crossed to a set of three spo11-1 mapping tester strains containing, among them, a recessive marker on each chromosome. The resulting spo11-1/spo11-1 diploids are sporulated and plated on drug-containing medium. Viable meiotic products that express the drug-resistance marker due to chromosome haploidization are selectively recovered. These meiotic products are haploid for most, but generally not all, chromosomes. The level of disomy for individual chromosomes averages 19%. Each of the recessive chromosomal markers is expressed in approximately a third of the drug-resistant segregants. Ninety-eight percent of these segregants show no evidence of intergenic recombination. Thus, two markers located on the same chromosome, but on different homologs, are virtually never expressed in the same drug-resistant clone. The utility of this mapping procedure is demonstrated by confirming the chromosomal location of seven known markers, as well as by the assignment of a previously unmapped mutation, spo12-1, to chromosome VIII. In addition, the analysis of the products of spo11-1 meiosis indicates that several markers previously assigned to either chromosome XIV or chromosome XVII are actually on the same chromosome.

EVIDENCE OF CHROMOSOMAL BREAKS NEAR THE MATING-TYPE LOCUS OF SACCHAROMYCES CEREVISIAE THAT ACCOMPANY MATα x MATα MATINGS

Genetics ◽  
1981 ◽  
Vol 99 (3-4) ◽  
pp. 383-403 ◽  
Author(s):  
John H McCusker ◽  
James E Haber
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Distal Portion ◽  
Chromosome Loss ◽  
Chromosome Break ◽  
Type Locus ◽  
Distal Marker ◽  
Chromosomal Breaks ◽  
A Cell ◽  
Chromosome Iii ◽  
The Right

ABSTRACT In order for two heterothallic MATα haploids of Saccharomyces cerevisiae to mate, one parent must apparently become, at least transiently, an a-like cell. Only about 25% of the matings result from an actual transposition of MATa sequences to replace MATα, and about 1% result from a deletion joining MAT to the normally silent HMRa allele. The majority of matings occur after an apparent chromosome break that deletes MATα and all of the known markers more distal on the right arm of chromosome III.——The chrdmosome break occurs at or very near MAT, invariably leaving the distal marker tsml hemizygous, but the closely linked proximal marker cry1 usually is heterozygous. The resulting diploid containing the broken chromosome is mitotically unstable; about 10% of the colonies contain visible sectors in which the rest of the broken chromosome is lost. The region close to the breakpoint (i.g., cryl) is unusually active in recombination. About 20% of the intact homologues remaining after chromosome loss were gene-converted for cryl. In addition, the broken end participated in reciprocal recombination events that joined the chromosome to the distal portion of the intact homologous chromosome.——The unstable diploids may also become stable and no longer give rise to mitotic segregants. We have found two distinct ways in which stabilization occurs. Most often the diploid becomes euploid by a recombination event that yields a cell homozygous for all markers distal to (and sometimes including) cryl. In one of 9 cases SO far analyzed, the stable diploid was still hemizygous for MATα and for other markers distal to MAT. This last case is similar to the healing of broken chromosomes in maize described by MCCLINTOCK(1 939,1941,1951).

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