Abstract
A method was developed for isolating large numbers of mutations on chromosome I of the yeast Saccharomyces cerevisiae. A strain monosomic for chromosome I (i.e., haploid for chromosome I and diploid for all other chromosomes) was mutagenized with either ethyl methanesulfonate or N-methyl-N'-nitro-N -nitrosoguanidine and screened for temperature-sensitive (Ts- ) mutants capable of growth on rich, glucose-containing medium at 25° but not at 37°. Recessive mutations induced on chromosome I are expressed, whereas those on the diploid chromosomes are usually not expressed because of the presence of wild-type alleles on the homologous chromosomes. Dominant ts mutations on all chromosomes should also be expressed, but these appeared rarely. — Of the 41 ts mutations analyzed, 32 mapped on chromosome I. These 32 mutations fell into only three complementation groups, which proved to be the previously described genes CDC15, CDC24 and PYK1 (or CDC19). We recovered 16 or 17 independent mutations in CDC15, 12 independent mutations in CDC24 and three independent mutations in PYK1. A fourth gene on chromosome I, MAK16, is known to be capable of giving rise to a ts-lethal allele, but we recovered no mutations in this gene. The remaining nine mutations isolated using the monosomic strain appeared not to map on chromosome I and were apparently expressed in the original mutants because they had become homozygous or hemizygous by mitotic recombination or chromosome loss. — The available information about the size of chromosome I suggests that it should contain approximately 60–100 genes. However, our isolation in the monosomic strain of multiple, independent alleles of just three genes suggests that only a small proportion of the genes on chromosome I is easily mutable to give a Ts--lethal phenotype. — During these studies, we located CDC24 on chromosome I and determined that it is centromere distal to PYK1 on the left arm of the chromosome.
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- Botstein D., Falco S. C., Stewart S. E., Brennan M., Scherer S., Stinchcomb D. T., Struhl K., Davis R. W. Sterile host yeasts (SHY): a eukaryotic system of biological containment for recombinant DNA experiments. Gene. 1979 Dec;8(1):17–24. doi: 10.1016/0378-1119(79)90004-0. [DOI] [PubMed] [Google Scholar]
- Brenner S. The genetics of Caenorhabditis elegans. Genetics. 1974 May;77(1):71–94. doi: 10.1093/genetics/77.1.71. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Broach J. R., Strathern J. N., Hicks J. B. Transformation in yeast: development of a hybrid cloning vector and isolation of the CAN1 gene. Gene. 1979 Dec;8(1):121–133. doi: 10.1016/0378-1119(79)90012-x. [DOI] [PubMed] [Google Scholar]
- Bruenn J., Mortimer R. K. Isolation of monosomics in yeast. J Bacteriol. 1970 May;102(2):548–551. doi: 10.1128/jb.102.2.548-551.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Burke R. L., Tekamp-Olson P., Najarian R. The isolation, characterization, and sequence of the pyruvate kinase gene of Saccharomyces cerevisiae. J Biol Chem. 1983 Feb 25;258(4):2193–2201. [PubMed] [Google Scholar]
- Cavalier-Smith T. Nuclear volume control by nucleoskeletal DNA, selection for cell volume and cell growth rate, and the solution of the DNA C-value paradox. J Cell Sci. 1978 Dec;34:247–278. doi: 10.1242/jcs.34.1.247. [DOI] [PubMed] [Google Scholar]
- Fried H. M., Pearson N. J., Kim C. H., Warner J. R. The genes for fifteen ribosomal proteins of Saccharomyces cerevisiae. J Biol Chem. 1981 Oct 10;256(19):10176–10183. [PubMed] [Google Scholar]
- Galau G. A., Klein W. H., Davis M. M., Wold B. J., Britten R. J., Davidson E. H. Structural gene sets active in embryos and adult tissues of the sea urchin. Cell. 1976 Apr;7(4):487–505. doi: 10.1016/0092-8674(76)90200-2. [DOI] [PubMed] [Google Scholar]
- Hartwell L. H., Mortimer R. K., Culotti J., Culotti M. Genetic Control of the Cell Division Cycle in Yeast: V. Genetic Analysis of cdc Mutants. Genetics. 1973 Jun;74(2):267–286. doi: 10.1093/genetics/74.2.267. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hereford L. M., Rosbash M. Number and distribution of polyadenylated RNA sequences in yeast. Cell. 1977 Mar;10(3):453–462. doi: 10.1016/0092-8674(77)90032-0. [DOI] [PubMed] [Google Scholar]
- Hereford L., Fahrner K., Woolford J., Jr, Rosbash M., Kaback D. B. Isolation of yeast histone genes H2A and H2B. Cell. 1979 Dec;18(4):1261–1271. doi: 10.1016/0092-8674(79)90237-x. [DOI] [PubMed] [Google Scholar]
- Holland J. P., Holland M. J. Structural comparison of two nontandemly repeated yeast glyceraldehyde-3-phosphate dehydrogenase genes. J Biol Chem. 1980 Mar 25;255(6):2596–2605. [PubMed] [Google Scholar]
- Ish-Horowicz D., Pinchin S. M., Schedl P., Artavanis-Tsakonas S., Mirault M. E. Genetic and molecular analysis of the 87A7 and 87C1 heat-inducible loci of D. melanogaster. Cell. 1979 Dec;18(4):1351–1358. doi: 10.1016/0092-8674(79)90245-9. [DOI] [PubMed] [Google Scholar]
- Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Judd B. H., Shen M. W., Kaufman T. C. The anatomy and function of a segment of the X chromosome of Drosophila melanogaster. Genetics. 1972 May;71(1):139–156. doi: 10.1093/genetics/71.1.139. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kaback D. B., Angerer L. M., Davidson N. Improved methods for the formation and stabilization of R-loops. Nucleic Acids Res. 1979 Jun 11;6(7):2499–2317. doi: 10.1093/nar/6.7.2499. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kaback D. B., Bhargava M. M., Halvorson H. O. Letter: Location and arrangement of genes coding for ribosomal RNA in Saccharomyces cerevisiae. J Mol Biol. 1973 Oct 5;79(4):735–739. doi: 10.1016/0022-2836(73)90076-4. [DOI] [PubMed] [Google Scholar]
- Kaback D. B., Halvorson H. O. Magnification of genes coding for ribosomal RNA in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1977 Mar;74(3):1177–1180. doi: 10.1073/pnas.74.3.1177. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kaback D. B., Halvorson H. O. Ribosomal DNA magnification in Saccharomyces cerevisiae. J Bacteriol. 1978 Apr;134(1):237–245. doi: 10.1128/jb.134.1.237-245.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kawasaki G., Fraenkel D. G. Cloning of yeast glycolysis genes by complementation. Biochem Biophys Res Commun. 1982 Oct 15;108(3):1107–1122. doi: 10.1016/0006-291x(82)92114-3. [DOI] [PubMed] [Google Scholar]
- Lauer G. D., Roberts T. M., Klotz L. C. Determination of the nuclear DNA content of Saccharomyces cerevisiae and implications for the organization of DNA in yeast chromosomes. J Mol Biol. 1977 Aug 25;114(4):507–526. doi: 10.1016/0022-2836(77)90175-9. [DOI] [PubMed] [Google Scholar]
- Lee A. S., Thomas T. L., Lev Z., Britten R. J., Davidson E. H. Four sizes of transcript produced by a single sea urchin gene expressed in early embryos. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3259–3263. doi: 10.1073/pnas.77.6.3259. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Luck D. J., Huang B., Piperno G. Genetic and biochemical analysis of the eukaryotic flagellum. Symp Soc Exp Biol. 1982;35:399–419. [PubMed] [Google Scholar]
- Mangiarotti G., Bozzaro S., Landfear S., Lodish H. F. Cell--cell contact, cyclic AMP, and gene expression during development of Dictyostelium discoideum. Curr Top Dev Biol. 1983;18:117–154. doi: 10.1016/s0070-2153(08)60581-5. [DOI] [PubMed] [Google Scholar]
- McAlister L., Holland M. J. Targeted deletion of a yeast enolase structural gene. Identification and isolation of yeast enolase isozymes. J Biol Chem. 1982 Jun 25;257(12):7181–7188. [PubMed] [Google Scholar]
- Moir D., Stewart S. E., Osmond B. C., Botstein D. Cold-sensitive cell-division-cycle mutants of yeast: isolation, properties, and pseudoreversion studies. Genetics. 1982 Apr;100(4):547–563. doi: 10.1093/genetics/100.4.547. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mortimer R. K., Schild D. Genetic map of Saccharomyces cerevisiae. Microbiol Rev. 1980 Dec;44(4):519–571. doi: 10.1128/mr.44.4.519-571.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nicklas J. A., Cline T. W. Vital Genes That Flank Sex-Lethal, an X-Linked Sex-Determining Gene of DROSOPHILA MELANOGASTER. Genetics. 1983 Apr;103(4):617–631. doi: 10.1093/genetics/103.4.617. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ouellette A. J. Purification by benzoylated cellulose chromatography of translatable messenger ribonucleic acid lacking polyadenylate. J Biol Chem. 1980 Apr 10;255(7):2740–2746. [PubMed] [Google Scholar]
- Reed S. I. The selection of S. cerevisiae mutants defective in the start event of cell division. Genetics. 1980 Jul;95(3):561–577. doi: 10.1093/genetics/95.3.561. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Regier J. C., Mazur G. D., Kafatos F. C. The silkmoth chorion: morphological and biochemical characterization of four surface regions. Dev Biol. 1980 May;76(2):286–304. doi: 10.1016/0012-1606(80)90380-2. [DOI] [PubMed] [Google Scholar]
- Ripoll P. Behavior of somatic cells homozygous for zygotic lethals in Drosophila melanogaster. Genetics. 1977 Jun;86(2 Pt 1):357–376. [PMC free article] [PubMed] [Google Scholar]
- Ripoll P., García-Bellido A. Viability of Homozygous Deficiencies in Somatic Cells of DROSOPHILA MELANOGASTER. Genetics. 1979 Mar;91(3):443–453. doi: 10.1093/genetics/91.3.443. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rogalski T. M., Moerman D. G., Baillie D. L. Essential genes and deficiencies in the unc-22 IV region of Caenorhabditis elegans. Genetics. 1982 Dec;102(4):725–736. doi: 10.1093/genetics/102.4.725. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rothstein R. J., Sherman F. Genes affecting the expression of cytochrome c in yeast: genetic mapping and genetic interactions. Genetics. 1980 Apr;94(4):871–889. doi: 10.1093/genetics/94.4.871. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shalit P., Loughney K., Olson M. V., Hall B. D. Physical analysis of the CYC1-sup4 interval in Saccharomyces cerevisiae. Mol Cell Biol. 1981 Mar;1(3):228–236. doi: 10.1128/mcb.1.3.228. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shortle D., Haber J. E., Botstein D. Lethal disruption of the yeast actin gene by integrative DNA transformation. Science. 1982 Jul 23;217(4557):371–373. doi: 10.1126/science.7046050. [DOI] [PubMed] [Google Scholar]
- Sina B. J., Pellegrini M. Genomic clones coding for some of the initial genes expressed during Drosophila development. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7351–7355. doi: 10.1073/pnas.79.23.7351. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sirotkin K., Davidson N. Developmentally regulated transcription from Drosophila melanogaster chromosomal site 67B. Dev Biol. 1982 Jan;89(1):196–210. doi: 10.1016/0012-1606(82)90307-4. [DOI] [PubMed] [Google Scholar]
- Sloat B. F., Adams A., Pringle J. R. Roles of the CDC24 gene product in cellular morphogenesis during the Saccharomyces cerevisiae cell cycle. J Cell Biol. 1981 Jun;89(3):395–405. doi: 10.1083/jcb.89.3.395. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith D. H., Berget P. B., King J. Temperature-sensitive mutants blocked in the folding or subunit assembly of the bacteriophage P22 tail-spike protein. I. Fine-structure mapping. Genetics. 1980 Oct;96(2):331–352. doi: 10.1093/genetics/96.2.331. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Snyder M., Davidson N. Two gene families clustered in a small region of the Drosophila genome. J Mol Biol. 1983 May 15;166(2):101–118. doi: 10.1016/s0022-2836(83)80001-1. [DOI] [PubMed] [Google Scholar]
- Spradling A. C., Rubin G. M. Drosophila genome organization: conserved and dynamic aspects. Annu Rev Genet. 1981;15:219–264. doi: 10.1146/annurev.ge.15.120181.001251. [DOI] [PubMed] [Google Scholar]
- St John T. P., Davis R. W. The organization and transcription of the galactose gene cluster of Saccharomyces. J Mol Biol. 1981 Oct 25;152(2):285–315. doi: 10.1016/0022-2836(81)90244-8. [DOI] [PubMed] [Google Scholar]
- Storti R. V., Szwast A. E. Molecular cloning and characterization of Drosophila genes and their expression during embryonic development and in primary muscle cell cultures. Dev Biol. 1982 Apr;90(2):272–283. doi: 10.1016/0012-1606(82)90376-1. [DOI] [PubMed] [Google Scholar]
- Tingle M. A., Küenzi M. T., Halvorson H. O. Germination of yeast spores lacking mitochondrial deoxyribonucleic acid. J Bacteriol. 1974 Jan;117(1):89–93. doi: 10.1128/jb.117.1.89-93.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Van Ness J., Maxwell I. H., Hahn W. E. Complex population of nonpolyadenylated messenger RNA in mouse brain. Cell. 1979 Dec;18(4):1341–1349. doi: 10.1016/0092-8674(79)90244-7. [DOI] [PubMed] [Google Scholar]
- Wickner R. B., Leibowitz M. J. Mak mutants of yeast: mapping and characterization. J Bacteriol. 1979 Oct;140(1):154–160. doi: 10.1128/jb.140.1.154-160.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wilkinson L. E., Pringle J. R. Transient G1 arrest of S. cerevisiae cells of mating type alpha by a factor produced by cells of mating type a. Exp Cell Res. 1974 Nov;89(1):175–187. doi: 10.1016/0014-4827(74)90200-6. [DOI] [PubMed] [Google Scholar]