Subcellular fluorescence localization analysis of all SAGA subunits in fission yeast (Schizosaccharomyces pombe)
Received date: 2013-07-16
Revised date: 2013-08-15
Online published: 2014-01-25
SAGA(Spt-Ada-Gcn5 Acetyltransferase complex) is a multi-subunit and conservative transcription complex, which is composed of 19 subunits in fission yeast and regulates the transcription of 10% genes in vivo. Through constructing in situ integrated fluorescence strains, we analyzed subcellular fluorescence localization of all SAGA subunits. Microscopic data showed localization manners could be sorted by 4 types, suggesting that these SAGA subunits may have additional functions besides transcriptional regulation. Subunit Sgf73 is the bridge that connects deubiquitination module and other SAGA modules, lacking of sgf73+ not only significantly reduced nuclear fluorescence localization (NFL) of deubiq-uitination subunits Ubp8, Sgf11, Sus1, but also affected NFL of acetylation subunits Gcn5, Sgf29, Ngg1, and the core structure subunit Spt7.The impact indicates that Sgf73 is important to maintain enzymatic function and stabilization of SAGA. Moreover, deletion of sgf73+ also caused a cytokinesis defect, which is characterized by a multi-nucleus and multi-septum phenotype. Overexpressing ace2+ and mid2+ in Dsgf73, which are key genes involved in septum degradation, showed that ace2+ could not rescue the defect, and mid2+ could only partially compensate for the deficiency, suggesting that Sgf73 may play a role in other pathways that affect cytokinesis.
Key words: SAGA; fluorescence localization; Sgf73; cytokinesis
Xing Zhou, Nan Zhou, Yao Yu, Hong Lv . Subcellular fluorescence localization analysis of all SAGA subunits in fission yeast (Schizosaccharomyces pombe)[J]. Hereditas(Beijing), 2014 , 36(2) : 169 -181 . DOI: 10.3724/SP.J.1005.2014.0169
[1] Huisinga KL, Pugh BF. A Genome-wide housekeeping role for TFIID and a highly regulated stress-related role for SAGA in Saccharomyces cerevisiae. Mol Cell, 2004, 13(4): 573–585. <\p>
[2] Zhang XY, Varthi M, Sykes SM, Phillips C, Warzecha C, Zhu WT, Wyce A, Thorne AW, Berger SL, Mcmahon SB. The putative cancer stem cell marker USP22 is a subunit of the human SAGA complex required for activated tran-scription and cell-cycle progression. Mol Cell, 2008, 29(1): 102–111. <\p>
[3] Koutelou E, Hirsch CL, Dent SY. Multiple faces of the SAGA complex. Curr Opin Cell Biol, 2010, 22(3): 374–382. <\p>
[4] Rodriguez-Navarro S. Insights into SAGA function during gene expression. EMBO Rep, 2009, 10(8): 843–850. <\p>
[5] Batta G, Szilagyi Z, Laczik M, Sipiczki M. The involvement of the Schizosaccharomyces pombe sep9/spt8 gene in the regulation of septum cleavage. FEMS Yeast Res, 2009, 9(5): 757–767. <\p>
[6] Helmlinger D, Marguerat S, Villén J, Gygi SP, Bähler J, Winston F. The S. pombe SAGA complex controls the switch from proliferation to sexual differentiation through the opposing roles of its subunits Gcn5 and Spt8. Genes Dev, 2008, 22(22): 3184–3195. <\p>
[7] Mcmahon SB, Wood MA, Cole MD. The essential cofactor TRRAP recruits the histone acetyltransferase hGCN5 to c-Myc. Mol Cell Biol, 2000, 20(2): 556–562. <\p>
[8] Rodńguez-Navarro S, Fischer T, Luo MJ, Antúnez O, Brettschneider S, Lechner J, Pérez-Ortín JE, Reed R, Hurt E. Sus1, a functional component of the SAGA histone acetylase complex and thes nuclear pore-associated mRNA export machinery. Cell, 2004, 116(1): 75–86. <\p>
[9] Hayashi A, Ding DQ, Tsutsumi C, Chikashige Y, Masuda H, Haraguchi T, Hiraoka Y. Localization of gene products using a chromosomally tagged GFP-fusion library in the fission yeast Schizosaccharomyces pombe. Genes Cells, 2009, 14(2): 217–225. <\p>
[10] Matsuyama A, Arai R, Yashiroda Y, Shirai A, Kamata A, Sekido S, Kobayashi Y, Hashimoto A, Hamamoto M, Hiraoka Y, Horinouchi S, Yoshida M. ORFeome cloning and global analysis of protein localization in the fission yeast Schizosac¬charomyces pombe. Nat Biotechnol, 2006, 24(7): 841–847. <\p>
[11] Helmlinger D, Hardy S, Sasorith S, Klein F, Robert F, Weber C, Miguet L, Potier N, Van-Dorsselaer A, Wurtz JM, Mandel JL, Tora L, Devys D. Ataxin-7 is a subunit of GCN5 histone acetyltransferase-containing complexes. Human Mol Genet, 2004, 13(12): 1257–1265. <\p>
[12] Köhler A, Schneider M, Cabal GG, Nehrbass U, Hurt E. Yeast Ataxin-7 links histone deubiquitination with gene gat-ing and mRNA export. Nat Cell Biol, 2008, 10(6): 707–715. <\p>
[13] Shukla A, Bajwa P, Bhaumik SR. SAGA-associated Sgf73p facilitates formation of the preinitiation complex assembly at the promoters either in a HAT-dependent or independent manner in vivo. Nucleic Acids Res, 2006, 34(21): 6225–6232. <\p>
[14] Palhan VB, Chen S, Peng GH, Tjernberg A, Gamper AM, Fan Y, Chait BT, La Spada AR, Roeder RG. Poly-glutamine-expanded ataxin-7 inhibits STAGA histone acetyltransferase activity to produce retinal degeneration. Proc Natl Acad Sci USA, 2005, 102(24): 8472–8477. <\p>
[15] Gregan J, Rabitsch PK, Rumpf C, Novatchkova M, Schleiffer A, Nasmyth K. High-throughput knockout screen in fission yeast. Nat Protoc, 2006, 1(5): 2457–2464. <\p>
[16] Grant PA, Schieltz D, Pray-Grant MG, Steger DJ, Reese JC, Yates JR, Workman JL. A subset of TAFIIs are integral components of the SAGA complex required for nucleosome acetylation and transcriptional stimulation. Cell, 1998, 94(1): 45–53. <\p>
[17] Huh WK, Falvo JV, Gerke LC, Carroll AS, Howson RW, Weissman JS, O'Shea EK. Global analysis of protein loca-lizatio
/
| 〈 |
|
〉 |