Abstract
When the fission yeastSchizosaccharomyces pombe is starved for nitrogen, the cells are arrested in the G1 phase, enter the G0 phase and initiate sexual development. Theste13 mutant, however, fails to undergo a G1 arrest when starved for nitrogen and since this mutant phenotype is not suppressed by a mutation in adenylyl cyclase (cyr1), it would appear thatste13 + either acts independently of the decrease in the cellular cAMP level induced by starvation for nitrogen, or functions downstream of this controlling event. We have used functional complementation to clone theste13 + gene from anS. pombe genomic library and show that its disruption is not lethal, indicating that, while the gene is required for sexual development, it is not essential for cell growth. Nucleotide sequencing predicts thatste13 + should encode a protein of 485 amino acids in which the consensus motifs of ATP-dependent RNA helicases of the DEAD box family are completely conserved. Point mutations introduced into these consensus motifs abolished theste13 + functions. The predicted Ste13 protein is 72% identical to theDrosophila melanogaster Me31B protein over a stretch of 391 amino acids. ME31B is a developmentally regulated gene that is expressed preferentially in the female germline and may be required for oogenesis. Expression of ME31B cDNA inS. pombe suppresses theste13 mutation. These two evolutionarily conserved genes encoding putative RNA helicases may play a pivotal role in sexual development.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Beach D, Nurse P (1981) High frequency transformation of the fission yeastSchizosaccharomyces pombe. Nature 290:140–142
Costello G, Rodgers L, Beach D (1986) Fission yeast enters the stationary phase G0 state from either mitotic G1 or G2. Curr Genet 11:119–125
De Valoir T, Tucker MA, Belikoff EJ, Camp LA, Bolduc C, Beckingham K (1991) A second maternally expressedDrosophila gene encodes a putative RNA helicase of the “DEAD box” family. Proc Natl Acad Sci USA 88:2113–2117
DeVoti J, Seydoux G, Beach D, McLeod M (1991) Interaction betweenranl + protein kinase and cAMP dependent protein kinase as negative regulators of fission yeast meiosis. EMBO J 10:3759–3768
Egel R (1971) Physiological aspects of conjugation in fission yeast. Planta 98:89–96
Egel R (1989) Mating-type genes, meiosis, and sporulation. In Molecular biology of the fission yeast, Nasim A, Young P, Johnson BF (eds), Academic Press, pp 31–73
Egel R, Egel-Mitani M (1974) Premeiotic DNA synthesis in fission yeast. Exp Cell Res 88:127–134
Engebrecht J, Voelkel-Meiman K, Roeder GS (1991) Meiosis-specific RNA splicing in yeast. Cell 66:1257–1268
Forsburg SL, Nurse P (1991) Identification of a G1-type cyclinpucl + in the fission yeastSchizosaccharomyces pombe. Nature 351:245–248
Girgsdies O (1982) Sterile mutants ofSchizosaccharomyces pombe; analysis by somatic hybridization. Curr Genet 6:223–227
Gutz H, Heslot H, Leupold L, Loprieno N (1974)Schizosaccharomyces pombe. In: King RC (ed) Handbook of genetics, vol 1. Plenum Press, New York, pp 395–446
Hay B, Jan LY, Jan YN (1988) A protein component ofDrosophila polar granules is encoded byvasa and has extensive sequence similarity to ATP-dependent helicases. Cell 55:577–587
Henikoff S (1984) Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28:351–359
Hereford L, Fahner K, Woolford J Jr, Rosbash M, Kabak DB (1979) Isolation of yeast histone genes H2A and H2B. Cell 18:1261–1271
Iggo RD, Lane DP (1989) Nuclear protein p68 is an RNA-dependent ATPase. EMBO J 8:1827–1831
Jensen R, Sprague GF Jr, Herskowitz I (1983) Regulation of yeast mating-type interconversion: Feedback control of HO gene expression by the mating-type locus. Proc Natl Acad Sci USA 80:3035–3039
Kiledjian M, Dreyfuss G (1992) Primary structure and binding activity of the hnRNP U protein: binding RNA through RGG box. EMBO J 11:2655–2664
Kishida M, Nagai T, Nakaseko Y, Shimoda C (1994) Meiosis-dependent mRNA splicing of the fission yeastSchizosaccharomyces pombe mes1 + gene. Curr Genet 25:497–503
Kitamura K, Shimoda C (1991) TheSchizosacharomyces pombe mam2 gene encodes a putative pheromone receptor which has a significant homology with theSaccharomyces cerevisiae Ste2 protein. EMBO J 10:3743–3751
Kitamura K, Nakagawa T, Shimoda C (1990) Novel sterile mutants of the fission yeastSchizosaccharomyces pombe which are defective in their response to starvation. Curr Genet 18:315–321
Lasko PF, Ashburner M (1988) The product of theDrosophila genevasa is very similar to eukaryotic initiation factor-4A. Nature 335:611–617
Leroy P, Alzari P, Sasson D, Wolgemuth D, Fellous M (1989) The protein encoded by murine male germ cell-specific transcript is a putative ATP-dependent RNA helicase. Cell 57:549–559
Leupold U, Sipiczki M (1991) Sterile UGA nonsense mutants of fission yeast. Curr Genet 20:67–73
Linder P, Slonimski PP (1989) An essential yeast protein, encoded by duplicated genesTIF1 andTIF2 and homologous to the mammalian translation initiation factor eIF-4A, can suppress a mitochondrial missense mutation. Proc Natl Acad Sci USA 86:2286–2290
Linder P, Lasko P, Leroy P, Nielsen P, Nishi K, Schnier J, Slonimski PP (1989) Birth of the D-E-A-D box. Nature 337:121–122
Lu, D Yunis JJ (1992) Cloning, expression and localization of an RNA helicase gene from a human lymphoid cell line with chromosomal breakpoint 11q23.3. Nucleic Acids Res 18:6485–6489
Maeda T, Motizuki N, Yamamoto M (1990) Adenylyl cyclase is dispensable for vegetative cell growth in the fission yeastSchizosaccharomyces pombe. Proc Natl Acad Sci USA 87:7814–7818
Michael H, Gutz H (1987) Sterility (ste) genes ofSchizosaccharomyces pombe. Yeast 3:5–9
Moreno S, Klar A, Nurse P (1991) Molecular genetic analysis of fission yeast. Methods Enzymol 194:795–823
Nielsen PJ, McMaster GK, Trachsel H (1985) Cloning of eukaryotic protein synthesis initiation factor genes: isolation and characterization of cDNA clones encoding factor eIF-4A. Nucleic Acids Res 13:6867–6880
Okazaki N, Okazaki K, Tanaka K, Okayama H (1991) Theste4 + gene, essential for sexual differentiation ofSchizosaccharomyces pombe, encodes a protein with a leucine zipper motif. Nucleic Acids Res 19:7043–7047
Pause A, Sonenberg N (1992) Mutational analysis of a DEAD box RNA helicase: the mammalian translation initiation factor eIF-4A. EMBO J 11:2643–2654
Rothstein R (1983) One step gene disruption in yeast. Methods Enzymol 101:202–211
Rozen F, Edery I, Meerovitch K, Dever TE, Merrick WC, Sonenberg N (1990) Bidirectional RNA helicase activity of eukaryotic translation initiation factors 4A and 4F. Mol Cell Biol 3:1134–1144
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New York
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Schmid SR, Linder P (1991) Translation initiation factor 4A fromSaccharomyces cerevisiae: Analysis of residues conserved in the D-E-A-D family of RNA helicases. Mol Cell Biol 11:3463–3471
Schmid SR, Linder P (1992) D-E-A-D protein family of putative RNA helicases. Mol Microbiol 6:283–292
Strahl-Bolsinger S, Tanner W (1993) A yeast gene encoding a putative RNA helicase of the “DEAD”-box family. Yeast 9:429–432
Thomas PS (1980) Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci USA 77:5201–5205
Thuriaux P, Sipiczki M, Fantes P (1980) Genetical analysis of a sterile mutant by protoplast fusion in the fission yeastSchizosaccharomyces pombe. J Gen Microbiol 116:525–528
Tommasino M, Maundrell K (1991) Uptake of thiamine bySchizosaccharomyces pombe and its effect as a transcriptional regulator of thiamine-sensitive genes. Curr Genet 20:63–66
Yanisch-Perron C, Vieira J, Messiong J (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33:103–119
Author information
Authors and Affiliations
Additional information
Communicated by K. Isono
Rights and permissions
About this article
Cite this article
Maekawa, H., Nakagawa, T., Uno, Y. et al. Theste13 + gene encoding a putative RNA helicase is essential for nitrogen starvation-induced G1 arrest and initiation of sexual development in the fission yeastSchizosaccharomyces pombe . Molec. Gen. Genet. 244, 456–464 (1994). https://doi.org/10.1007/BF00583896
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00583896