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      A Functional Link Between Bir1 and the Saccharomyces cerevisiae Ctf19 Kinetochore Complex Revealed Through Quantitative Fitness Analysis

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          Abstract

          The chromosomal passenger complex (CPC) is a key regulator of eukaryotic cell division, consisting of the protein kinase Aurora B/ Ipl1 in association with its activator (INCENP/ Sli15) and two additional proteins (Survivin/ Bir1 and Borealin/ Nbl1). Here, we report a genome-wide genetic interaction screen in Saccharomyces cerevisiae using the bir1-17 mutant, identifying through quantitative fitness analysis deletion mutations that act as enhancers and suppressors. Gene knockouts affecting the Ctf19 kinetochore complex were identified as the strongest enhancers of bir1-17 , while mutations affecting the large ribosomal subunit or the mRNA nonsense-mediated decay pathway caused strong phenotypic suppression. Thus, cells lacking a functional Ctf19 complex become highly dependent on Bir1 function and vice versa. The negative genetic interaction profiles of bir1-17 and the cohesin mutant mcd1-1 showed considerable overlap, underlining the strong functional connection between sister chromatid cohesion and chromosome biorientation. Loss of some Ctf19 components, such as Iml3 or Chl4, impacted differentially on bir1-17 compared with mutations affecting other CPC components: despite the synthetic lethality shown by either iml3 ∆ or chl4 ∆ in combination with bir1-17 , neither gene knockout showed any genetic interaction with either ipl1-321 or sli15-3 . Our data therefore imply a specific functional connection between the Ctf19 complex and Bir1 that is not shared with Ipl1.

          Most cited references63

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          Three new dominant drug resistance cassettes for gene disruption in Saccharomyces cerevisiae.

          Disruption-deletion cassettes are powerful tools used to study gene function in many organisms, including Saccharomyces cerevisiae. Perhaps the most widely useful of these are the heterologous dominant drug resistance cassettes, which use antibiotic resistance genes from bacteria and fungi as selectable markers. We have created three new dominant drug resistance cassettes by replacing the kanamycin resistance (kan(r)) open reading frame from the kanMX3 and kanMX4 disruption-deletion cassettes (Wach et al., 1994) with open reading frames conferring resistance to the antibiotics hygromycin B (hph), nourseothricin (nat) and bialaphos (pat). The new cassettes, pAG25 (natMX4), pAG29 (patMX4), pAG31 (patMX3), pAG32 (hphMX4), pAG34 (hphMX3) and pAG35 (natMX3), are cloned into pFA6, and so are in all other respects identical to pFA6-kanMX3 and pFA6-kanMX4. Most tools and techniques used with the kanMX plasmids can also be used with the hph, nat and patMX containing plasmids. These new heterologous dominant drug resistance cassettes have unique antibiotic resistance phenotypes and do not affect growth when inserted into the ho locus. These attributes make the cassettes ideally suited for creating S. cerevisiae strains with multiple mutations within a single strain. Copyright 1999 John Wiley & Sons, Ltd.
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            Improved method for high efficiency transformation of intact yeast cells.

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              Cohesin: its roles and mechanisms.

              The cohesin complex is a major constituent of interphase and mitotic chromosomes. Apart from its role in mediating sister chromatid cohesion, it is also important for DNA double-strand-break repair and transcriptional control. The functions of cohesin are regulated by phosphorylation, acetylation, ATP hydrolysis, and site-specific proteolysis. Recent evidence suggests that cohesin acts as a novel topological device that traps chromosomal DNA within a large tripartite ring formed by its core subunits.
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                Author and article information

                Journal
                G3 (Bethesda)
                Genetics
                G3: Genes, Genomes, Genetics
                G3: Genes, Genomes, Genetics
                G3: Genes, Genomes, Genetics
                G3: Genes|Genomes|Genetics
                Genetics Society of America
                2160-1836
                28 July 2017
                September 2017
                : 7
                : 9
                : 3203-3215
                Affiliations
                [* ]Centre for Gene Regulation and Expression, University of Dundee, DD1 5EH, UK
                []Wellcome Trust Centre for Cell Biology, University of Edinburgh, EH9 3BF, UK
                []Institute for Cell and Molecular Biosciences, Newcastle University, NE2 4HH, UK
                Author notes
                [1]

                Present address: Wellcome Trust Centre for Cell Biology, Michael Swann Building, The King’s Buildings, University of Edinburgh, Max Born Crescent, Edinburgh EH9 3BF, Scotland, UK.

                [2]

                Present address: Institute of Bioorganic Chemistry PAN, ul. Z. Noskowskiego 12/14, 61-704 Poznań, Poland.

                [3]

                Present address: Division of Microbiology and Molecular Medicine, Department of Clinical and Experimental Medicine, Linköping University, SE-581 83 Linköping, Sweden.

                [4 ]Corresponding author: Centre for Gene Regulation and Expression, School of Life Sciences, MSI/WTB Complex, University of Dundee, Dow St., Dundee DD1 5EH, Scotland. E-mail: m.j.r.stark@ 123456dundee.ac.uk
                Author information
                http://orcid.org/0000-0003-0668-6157
                http://orcid.org/0000-0002-4186-8506
                http://orcid.org/0000-0002-9582-4242
                http://orcid.org/0000-0002-3596-9407
                http://orcid.org/0000-0003-2478-085X
                http://orcid.org/0000-0001-9086-191X
                Article
                GGG_300089
                10.1534/g3.117.300089
                5592945
                28754723
                8b6629f5-c71a-4420-9f1a-299de830a891
                Copyright © 2017 Makrantoni et al.

                This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 14 May 2017
                : 25 July 2017
                Page count
                Figures: 5, Tables: 2, Equations: 0, References: 75, Pages: 13
                Categories
                Investigations

                Genetics
                bir1,chromosome biorientation,kinetochore,iml3-chl4 complex,yeast
                Genetics
                bir1, chromosome biorientation, kinetochore, iml3-chl4 complex, yeast

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