title: A role for the budding yeast separase, Esp1, in Ty1 element retrotransposition.
abstract: Separase/Esp1 is a protease required at the onset of anaphase to cleave cohesin and thereby enable sister chromatid separation. Esp1 also promotes release of the Cdc14 phosphatase from the nucleolus to enable mitotic exit. To uncover other potential roles for separase, we performed two complementary genome-wide genetic interaction screens with a strain carrying the budding yeast esp1-1 separase mutation. We identified 161 genes that when mutated aggravate esp1-1 growth and 44 genes that upon increased dosage are detrimental to esp1-1 viability. In addition to the expected cell cycle and sister chromatid segregation genes that were identified, 24% of the genes identified in the esp1-1 genetic screens have a role in Ty1 element retrotransposition. Retrotransposons, like retroviruses, replicate through reverse transcription of an mRNA intermediate and the resultant cDNA product is integrated into the genome by a conserved transposon or retrovirus encoded integrase protein. We purified Esp1 from yeast and identified an interaction between Esp1 and Ty1 integrase using mass spectrometry that was subsequently confirmed by co-immunoprecipitation analysis. Ty1 transposon mobility and insertion upstream of the SUF16 tRNA gene are both reduced in an esp1-1 strain but increased in cohesin mutant strains. Securin/Pds1, which is required for efficient localization of Esp1 to the nucleus, is also required for efficient Ty1 transposition. We propose that Esp1 serves two roles to mediate Ty1 transposition - one to remove cohesin and the second to target Ty1-IN to chromatin.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia, Canada; Wine Research Centre, University of British Columbia, Vancouver, British Columbia, Canada.
authors.forename: Krystina L
authors.initials: KL
authors.lastname: Ho
authors.affiliation: Wine Research Centre, University of British Columbia, Vancouver, British Columbia, Canada.
authors.forename: Lina
authors.initials: L
authors.lastname: Ma
authors.affiliation: Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia, Canada; Wine Research Centre, University of British Columbia, Vancouver, British Columbia, Canada.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Cheung
authors.affiliation: Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia, Canada; Wine Research Centre, University of British Columbia, Vancouver, British Columbia, Canada.
authors.forename: Savrina
authors.initials: S
authors.lastname: Manhas
authors.affiliation: Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia, Canada; Centre for High-Throughput Biology, University of British Columbia, Vancouver, British Columbia, Canada.
authors.forename: Nancy
authors.initials: N
authors.lastname: Fang
authors.affiliation: Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia, Canada; Wine Research Centre, University of British Columbia, Vancouver, British Columbia, Canada.
authors.forename: Kaiqian
authors.initials: K
authors.lastname: Wang
authors.affiliation: Department of Cellular and Physiological Sciences, University of British Columbia, Vancouver, British Columbia, Canada.
authors.forename: Barry
authors.initials: B
authors.lastname: Young
authors.affiliation: Department of Cellular and Physiological Sciences, University of British Columbia, Vancouver, British Columbia, Canada.
authors.forename: Christopher
authors.initials: C
authors.lastname: Loewen
authors.affiliation: Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia, Canada; Centre for High-Throughput Biology, University of British Columbia, Vancouver, British Columbia, Canada.
authors.forename: Thibault
authors.initials: T
authors.lastname: Mayor
authors.affiliation: Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia, Canada; Wine Research Centre, University of British Columbia, Vancouver, British Columbia, Canada.
authors.forename: Vivien
authors.initials: V
authors.lastname: Measday
mesh_terms: D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D002466:Cell Nucleolus / Q000235:genetics; D002843:Chromatin / Q000235:genetics* / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism; D020090:Chromosome Segregation / Q000235:genetics*; D018076:DNA, Complementary; D008938:Mitosis / Q000235:genetics*; D012343:RNA, Transfer / Q000235:genetics; D018626:Retroelements / Q000235:genetics*; D012441:Saccharomyces cerevisiae; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism; D064249:Securin / Q000235:genetics / Q000378:metabolism; D064247:Separase / Q000235:genetics* / Q000378:metabolism; D000097722:Cohesins
chemical_list: D018797:Cell Cycle Proteins; D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D018076:DNA, Complementary; D012343:RNA, Transfer; D018626:Retroelements; D029701:Saccharomyces cerevisiae Proteins; D064249:Securin; D064247:Separase; D000097722:Cohesins; C578004:PDS1 protein, S cerevisiae; C577986:ESP1 protein, S cerevisiae
references.citation: Dev Cell. 2009 Sep;17(3):344-54
references.pmid: 19758559
references.citation: Genetics. 2008 Jan;178(1):197-214
references.pmid: 18202368
references.citation: Nature. 2004 Jul 29;430(6999):573-8
references.pmid: 15229615
references.citation: Genetics. 1988 Feb;118(2):203-12
references.pmid: 2834263
references.citation: Cell. 2000 Oct 27;103(3):375-86
references.pmid: 11081625
references.citation: Cell. 2002 Jan 25;108(2):207-20
references.pmid: 11832211
references.citation: Genetics. 1998 Apr;148(4):1743-61
references.pmid: 9560391
references.citation: BMC Bioinformatics. 2013;14:354
references.pmid: 24305553
references.citation: Nat Cell Biol. 2001 Sep;3(9):771-7
references.pmid: 11533655
references.citation: Cell. 2001 May 18;105(4):459-72
references.pmid: 11371343
references.citation: J Cell Biol. 2014 Oct 27;207(2):189-99
references.pmid: 25332162
references.citation: Genetics. 2000 Feb;154(2):543-56
references.pmid: 10655210
references.citation: Nat Rev Mol Cell Biol. 2001 Nov;2(11):815-26
references.pmid: 11715048
references.citation: Nat Protoc. 2007;2(8):1896-906
references.pmid: 17703201
references.citation: Science. 2001 Dec 14;294(5550):2364-8
references.pmid: 11743205
references.citation: Mol Cell Biol. 1988 Dec;8(12):5386-97
references.pmid: 3072479
references.citation: Mol Cell Biol. 2001 Aug;21(16):5374-88
references.pmid: 11463820
references.citation: Mol Cell Biol. 2001 Oct;21(19):6606-14
references.pmid: 11533248
references.citation: Genes Dev. 2005 Apr 15;19(8):955-64
references.pmid: 15833918
references.citation: PLoS One. 2011;6(4):e19459
references.pmid: 21559383
references.citation: J Proteome Res. 2006 Apr;5(4):988-94
references.pmid: 16602707
references.citation: Genes Dev. 2008 Aug 15;22(16):2215-27
references.pmid: 18708580
references.citation: Genes Dev. 1996 Dec 15;10(24):3081-93
references.pmid: 8985178
references.citation: Mol Cell Biol. 1988 May;8(5):2159-65
references.pmid: 2455217
references.citation: Genes Dev. 2005 Apr 15;19(8):942-54
references.pmid: 15833917
references.citation: PLoS Genet. 2012;8(8):e1002856
references.pmid: 22912589
references.citation: Annu Rev Genet. 2009;43:525-58
references.pmid: 19886810
references.citation: Genome Res. 2003 Nov;13(11):2498-504
references.pmid: 14597658
references.citation: Genetics. 2003 Jan;163(1):55-67
references.pmid: 12586696
references.citation: Genetics. 2006 Apr;172(4):2157-67
references.pmid: 16415356
references.citation: Genome Res. 2012 Apr;22(4):693-703
references.pmid: 22219510
references.citation: Mol Cell. 2009 Mar 27;33(6):763-74
references.pmid: 19328069
references.citation: Nat Cell Biol. 2003 Mar;5(3):249-54
references.pmid: 12598903
references.citation: Mol Cell Biol. 1991 Nov;11(11):5710-7
references.pmid: 1922073
references.citation: Science. 2014 Nov 21;346(6212):963-7
references.pmid: 25414305
references.citation: J Cell Biol. 1996 Apr;133(1):85-97
references.pmid: 8601616
references.citation: Cell. 1984 Dec;39(3 Pt 2):675-82
references.pmid: 6096019
references.citation: Yeast. 1994 Jan;10(1):81-92
references.pmid: 8203154
references.citation: Science. 2004 Feb 6;303(5659):808-13
references.pmid: 14764870
references.citation: Science. 2003 Nov 21;302(5649):1399-401
references.pmid: 14631041
references.citation: Cell. 1993 Jun 4;73(5):1007-18
references.pmid: 8388781
references.citation: Mol Cell. 2006 Feb 3;21(3):319-30
references.pmid: 16455487
references.citation: Bioinformatics. 2009 Apr 15;25(8):1091-3
references.pmid: 19237447
references.citation: Annu Rev Genet. 2008;42:587-617
references.pmid: 18680436
references.citation: Genes Dev. 1999 Feb 1;13(3):320-33
references.pmid: 9990856
references.citation: Genetics. 2004 Nov;168(3):1159-76
references.pmid: 15579677
references.citation: Mob DNA. 2012 Aug 02;3(1):12
references.pmid: 22856544
references.citation: Mol Cell. 1999 Sep;4(3):445-50
references.pmid: 10518226
references.citation: Mol Cell Biol. 1998 Feb;18(2):1105-14
references.pmid: 9448008
references.citation: Cell Cycle. 2004 Feb;3(2):182-8
references.pmid: 14712087
references.citation: Genetics. 2001 Dec;159(4):1449-65
references.pmid: 11779788
references.citation: Nat Struct Mol Biol. 2005 Jun;12(6):552-3
references.pmid: 15880121
references.citation: Yeast. 1999 Oct;15(14):1541-53
references.pmid: 10514571
references.citation: Nucleic Acids Res. 1998 Feb 1;26(3):689-96
references.pmid: 9443958
references.citation: Cell. 1997 Oct 3;91(1):35-45
references.pmid: 9335333
references.citation: Nature. 2013 Jan 10;493(7431):250-4
references.pmid: 23178808
references.citation: Science. 2014 Apr 4;344(6179):55-8
references.pmid: 24674868
references.citation: Cytogenet Genome Res. 2005;110(1-4):70-90
references.pmid: 16093660
references.citation: Genome Res. 2012 Apr;22(4):704-13
references.pmid: 22219511
references.citation: Mol Cell Biol. 1992 Jun;12(6):2813-25
references.pmid: 1317008
references.citation: J Cell Biol. 2001 Jan 8;152(1):27-40
references.pmid: 11149918
references.citation: Annu Rev Genet. 2004;38:203-32
references.pmid: 15568976
references.citation: Cell. 2004 Jul 9;118(1):31-44
references.pmid: 15242642
references.citation: Mol Cell Biol. 2000 Apr;20(7):2436-45
references.pmid: 10713167
references.citation: Cell. 2001 Dec 28;107(7):929-39
references.pmid: 11779468
references.citation: Mol Cell Biol. 2010 Jan;30(2):382-98
references.pmid: 19901074
references.citation: Cell. 1999 Jul 23;98(2):249-59
references.pmid: 10428036
references.citation: Mol Cell Proteomics. 2005 Dec;4(12):2010-21
references.pmid: 16249172
references.citation: Nucleic Acids Res. 2008 Aug;36(13):4317-26
references.pmid: 18586821
references.citation: Nature. 2002 Jul 25;418(6896):387-91
references.pmid: 12140549
references.citation: Genetics. 1984 Jun;107(2):179-97
references.pmid: 6329902
references.citation: Genome Res. 2005 May;15(5):641-54
references.pmid: 15837808
references.citation: Genetics. 2004 Nov;168(3):1763-71
references.pmid: 15579722
references.citation: Proc Natl Acad Sci U S A. 2005 Sep 27;102(39):13956-61
references.pmid: 16172405
references.citation: Anal Chem. 2003 Feb 1;75(3):663-70
references.pmid: 12585499
references.citation: Cell. 1988 Sep 23;54(7):955-66
references.pmid: 2843295
references.citation: Nat Genet. 2014 Oct;46(10):1147-51
references.pmid: 25173104
references.citation: Genes Dev. 1988 Feb;2(2):150-9
references.pmid: 2834270
references.citation: Mol Cell. 2000 Feb;5(2):243-54
references.pmid: 10882066
references.citation: Proc Natl Acad Sci U S A. 1991 Feb 1;88(3):936-40
references.pmid: 1846969
references.citation: Methods Mol Biol. 2006;313:171-92
references.pmid: 16118434
references.citation: Methods Enzymol. 2002;350:316-26
references.pmid: 12073321
references.citation: Curr Biol. 2003 Feb 4;13(3):R104-14
references.pmid: 12573239
references.citation: Mol Cell Biol. 1998 May;18(5):2502-13
references.pmid: 9566871
references.citation: Genome Res. 1998 May;8(5):464-78
references.pmid: 9582191
references.citation: Mol Biol Cell. 2009 Mar;20(5):1576-91
references.pmid: 19144818
references.citation: Genetics. 2008 Apr;178(4):2361-72
references.pmid: 18430955
references.citation: J Cell Sci. 2005 Feb 15;118(Pt 4):733-42
references.pmid: 15671062
references.citation: Cell Cycle. 2004 Jul;3(7):960-7
references.pmid: 15190202
references.citation: Genetics. 2003 Jul;164(3):867-79
references.pmid: 12871900
references.citation: Nature. 1999 Jul 1;400(6739):37-42
references.pmid: 10403247
references.citation: Proc Natl Acad Sci U S A. 2005 Nov 29;102(48):17308-13
references.pmid: 16260736
references.citation: Genes Dev. 1996 Mar 1;10(5):620-33
references.pmid: 8598291
references.citation: Cell Cycle. 2006 Mar;5(6):592-5
references.pmid: 16582600
references.citation: BMC Mol Biol. 2009;10:99
references.pmid: 19874630
references.citation: Nature. 2004 Aug 26;430(7003):1044-8
references.pmid: 15329725
references.citation: J Biol Chem. 2012 May 25;287(22):18589-95
references.pmid: 22493285
references.citation: Mol Cell Biol. 1998 Feb;18(2):1115-24
references.pmid: 9448009
references.citation: Eur J Biochem. 1999 Jul;263(1):14-9
references.pmid: 10429181
references.citation: Methods Mol Biol. 2000;146:1-16
references.pmid: 10948493
references.citation: Genes Dev. 2004 Nov 1;18(21):2581-95
references.pmid: 15520278
references.citation: EMBO J. 2004 Aug 4;23(15):3144-53
references.pmid: 15241476