C. elegans PAT-9 is a nuclear zinc finger protein critical for the assembly of muscle attachments

Cell & Bioscience

Peter L Jones²  Benjamin D Williams¹  Lauren Rogowski¹  Mitchell Meghpara¹  Rebecca A Bachmann¹  Takako I Jones²  Qian Liu¹ 

[1] Department of Cell and Developmental Biology, University of Illinois at Urbana-Champaign, 601 S. Goodwin Ave, B107 Chemical and Life Sciences Laboratory, Urbana, IL, 61801, USA;Present Address: Boston Biomedical Research Institute, 64 Grove St., Watertown, MA, 02472, USA

关键词: Pat;    Zinc finger;    Muscle;    Sarcomere;   

Others  :  793471
DOI  :  10.1186/2045-3701-2-18

 received in 2012-04-17, accepted in 2012-05-22,  发布年份 2012

【 摘 要 】

Background

Caenorhabditis elegans sarcomeres have been studied extensively utilizing both forward and reverse genetic techniques to provide insight into muscle development and the mechanisms behind muscle contraction. A previous genetic screen investigating early muscle development produced 13 independent mutant genes exhibiting a Pat (

p

aralyzed and

a

rrested elongation at the

t

wo-fold length of embryonic development) muscle phenotype. This study reports the identification and characterization of one of those genes, pat-9.

Results

Positional cloning, reverse genetics, and plasmid rescue experiments were used to identify the predicted C. elegans gene T27B1.2 (recently named ztf-19) as the pat-9 gene. Analysis of pat-9 showed it is expressed early in development and within body wall muscle lineages, consistent with a role in muscle development and producing a Pat phenotype. However, unlike most of the other known Pat gene family members, which encode structural components of muscle attachment sites, PAT-9 is an exclusively nuclear protein. Analysis of the predicted PAT-9 amino acid sequence identified one putative nuclear localization domain and three C2H2 zinc finger domains. Both immunocytochemistry and PAT-9::GFP fusion expression confirm that PAT-9 is primarily a nuclear protein and chromatin immunoprecipitation (ChIP) experiments showed that PAT-9 is present on certain gene promoters.

Conclusions

We have shown that the T27B1.2 gene is pat-9. Considering the Pat-9 mutant phenotype shows severely disrupted muscle attachment sites despite PAT-9 being a nuclear zinc finger protein and not a structural component of muscle attachment sites, we propose that PAT-9 likely functions in the regulation of gene expression for some necessary structural or regulatory component(s) of the muscle attachment sites.

【 授权许可】

   
2012 Liu et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

【 参考文献 】

• [1]Moerman DG, Williams BD: Sarcomere assembly in C. elegans muscle. WormBook 2006, 1-16.
• [2]Qadota H, Benian GM: Molecular structure of sarcomere-to-membrane attachment at M-Lines in C. elegans muscle. J Biomed Biotechnol 2010, 2010:864749.
• [3]Mackenzie JM, Garcea RL, Zengel JM, Epstein HF: Muscle development in Caenorhabditis elegans: mutants exhibiting retarded sarcomere construction. Cell 1978, 15:751-762.
• [4]Francis GR, Waterston RH: Muscle organization in Caenorhabditis elegans: localization of proteins implicated in thin filament attachment and I-band organization. J Cell Biol 1985, 101:1532-1549.
• [5]Hresko MC, Williams BD, Waterston RH: Assembly of body wall muscle and muscle cell attachment structures in Caenorhabditis elegans. J Cell Biol 1994, 124:491-506.
• [6]Liu Q, Jones TI, Tang VW, Brieher WM, Jones PL: Facioscapulohumeral muscular dystrophy region gene-1 (FRG-1) is an actin bundling protein associated with muscle attachment sites. J Cell Sci 2010, 123:1116-1123.
• [7]Lecroisey C, Segalat L, Gieseler K: The C. elegans dense body: anchoring and signaling structure of the muscle. J Muscle Res Cell Motil 2007, 28:79-87.
• [8]Hanel ML, Sun CY, Jones TI, Long SW, Zanotti S, Milner D, Jones PL: Facioscapulohumeral muscular dystrophy (FSHD) region gene 1 (FRG1) is a dynamic nuclear and sarcomeric protein. Differentiation 2010, 2:107-118.
• [9]Barstead RJ, Waterston RH: The basal component of the nematode dense-body is vinculin. J Biol Chem 1989, 264:10177-10185.
• [10]Williams BD, Waterston RH: Genes critical for muscle development and function in Caenorhabditis elegans identified through lethal mutations. J Cell Biol 1994, 124:475-490.
• [11]Waterston RH, Thomson JN, Brenner S: Mutants with altered muscle structure of Caenorhabditis elegans. Dev Biol 1980, 77:271-302.
• [12]Lecroisey C, Martin E, Mariol MC, Granger L, Schwab Y, Labouesse M, Segalat L, Gieseler K: DYC-1, a protein functionally linked to dystrophin in Caenorhabditis elegans is associated with the dense body, where it interacts with the muscle LIM domain protein ZYX-1. Mol Biol Cell 2008, 19:785-796.
• [13]Broday L, Kolotuev I, Didier C, Bhoumik A, Podbilewicz B, Ronai Z: The LIM domain protein UNC-95 is required for the assembly of muscle attachment structures and is regulated by the RING finger protein RNF-5 in C. elegans. J Cell Biol 2004, 165:857-867.
• [14]Hobert O, Moerman DG, Clark KA, Beckerle MC, Ruvkun G: A conserved LIM protein that affects muscular adherens junction integrity and mechanosensory function in Caenorhabditis elegans. J Cell Biol 1999, 144:45-57.
• [15]Mercer KB, Flaherty DB, Miller RK, Qadota H, Tinley TL, Moerman DG, Benian GM: Caenorhabditis elegans UNC-98, a C2H2 Zn finger protein, is a novel partner of UNC-97/PINCH in muscle adhesion complexes. Mol Biol Cell 2003, 14:2492-2507.
• [16]Mackinnon AC, Qadota H, Norman KR, Moerman DG, Williams BD: C. elegans PAT-4/ILK functions as an adaptor protein within integrin adhesion complexes. Curr Biol 2002, 12:787-797.
• [17]Lin X, Qadota H, Moerman DG, Williams BD: C. elegans PAT-6/actopaxin plays a critical role in the assembly of integrin adhesion complexes in vivo. Curr Biol 2003, 13:922-932.
• [18]Norman KR, Cordes S, Qadota H, Rahmani P, Moerman DG: UNC-97/PINCH is involved in the assembly of integrin cell adhesion complexes in Caenorhabditis elegans body wall muscle. Dev Biol 2007, 309:45-55.
• [19]Williams BD, Schrank B, Huynh C, Shownkeen R, Waterston RH: A genetic mapping system in Caenorhabditis elegans based on polymorphic sequence-tagged sites. Genetics 1992, 131:609-624.
• [20]Meissner B, Warner A, Wong K, Dube N, Lorch A, McKay SJ, Khattra J, Rogalski T, Somasiri A, Chaudhry I, et al.: An integrated strategy to study muscle development and myofilament structure in Caenorhabditis elegans. PLoS Genet 2009, 5:e1000537.
• [21]Marfori M, Mynott A, Ellis JJ, Mehdi AM, Saunders NF, Curmi PM, Forwood JK, Boden M, Kobe B: Molecular basis for specificity of nuclear import and prediction of nuclear localization. Biochim Biophys Acta 2010, 1813:1562-1577.
• [22]Barrasa MI, Vaglio P, Cavasino F, Jacotot L, Walhout AJ: EDGEdb: a transcription factor-DNA interaction database for the analysis of C. elegans differential gene expression. BMC Genomics 2007, 8:21. BioMed Central Full Text
• [23]Iuchi S: Three classes of C2H2 zinc finger proteins. Cell Mol Life Sci 2001, 58:625-635.
• [24]Lakowski B, Eimer S, Gobel C, Bottcher A, Wagler B, Baumeister R: Two suppressors of sel-12 encode C2H2 zinc-finger proteins that regulate presenilin transcription in Caenorhabditis elegans. Development 2003, 130:2117-2128.
• [25]Wang Y, Gracheva EO, Richmond J, Kawano T, Couto JM, Calarco JA, Vijayaratnam V, Jin Y, Zhen M: The C2H2 zinc-finger protein SYD-9 is a putative posttranscriptional regulator for synaptic transmission. Proc Natl Acad Sci U S A 2006, 103:10450-10455.
• [26]Roy Chowdhuri S, Crum T, Woollard A, Aslam S, Okkema PG: The T-box factor TBX-2 and the SUMO conjugating enzyme UBC-9 are required for ABa-derived pharyngeal muscle in C. elegans. Dev Biol 2006, 295:664-677.
• [27]Thatcher JD, Haun C, Okkema PG: The DAF-3 Smad binds DNA and represses gene expression in the Caenorhabditis elegans pharynx. Development 1999, 126:97-107.
• [28]Patterson GI, Koweek A, Wong A, Liu Y, Ruvkun G: The DAF-3 Smad protein antagonizes TGF-beta-related receptor signaling in the Caenorhabditis elegans dauer pathway. Genes Dev 1997, 11:2679-2690.
• [29]Sonnichsen B, Koski LB, Walsh A, Marschall P, Neumann B, Brehm M, Alleaume AM, Artelt J, Bettencourt P, Cassin E, et al.: Full-genome RNAi profiling of early embryogenesis in Caenorhabditis elegans. Nature 2005, 434:462-469.
• [30]Simmer F, Moorman C, van der Linden AM, Kuijk E, van den Berghe PV, Kamath RS, Fraser AG, Ahringer J, Plasterk RH: Genome-wide RNAi of C. elegans using the hypersensitive rrf-3 strain reveals novel gene functions. PLoS Biol 2003, 1:E12.
• [31]Vowels JJ, Thomas JH: Genetic analysis of chemosensory control of dauer formation in Caenorhabditis elegans. Genetics 1992, 130:105-123.
• [32]Palmer RE, Inoue T, Sherwood DR, Jiang LI, Sternberg PW: Caenorhabditis elegans cog-1 locus encodes GTX/Nkx6.1 homeodomain proteins and regulates multiple aspects of reproductive system development. Dev Biol 2002, 252:202-213.
• [33]Reinhart BJ, Slack FJ, Basson M, Pasquinelli AE, Bettinger JC, Rougvie AE, Horvitz HR, Ruvkun G: The 21-nucleotide let-7 RNA regulates developmental timing in Caenorhabditis elegans. Nature 2000, 403:901-906.
• [34]Martinez NJ, Ow MC, Reece-Hoyes JS, Barrasa MI, Ambros VR, Walhout AJ: Genome-scale spatiotemporal analysis of Caenorhabditis elegans microRNA promoter activity. Genome Res 2008, 18:2005-2015.
• [35]Brenner S: The genetics of Caenorhabditis elegans. Genetics 1974, 77:71-94.
• [36]Wicks SR, Yeh RT, Gish WR, Waterston RH, Plasterk RH: Rapid gene mapping in Caenorhabditis elegans using a high density polymorphism map. Nat Genet 2001, 28:160-164.
• [37]Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE, Mello CC: Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 1998, 391:806-811.
• [38]Timmons L, Court DL, Fire A: Ingestion of bacterially expressed dsRNAs can produce specific and potent genetic interference in Caenorhabditis elegans. Gene 2001, 263:103-112.
• [39]Mukhopadhyay A, Deplancke B, Walhout AJ, Tissenbaum HA: Chromatin immunoprecipitation (ChIP) coupled to detection by quantitative real-time PCR to study transcription factor binding to DNA in Caenorhabditis elegans. Nat Protoc 2008, 3:698-709.
• [40]Barstead RJ, Kleiman L, Waterston RH: Cloning, sequencing, and mapping of an alpha-actinin gene from the nematode Caenorhabditis elegans. Cell Motil Cytoskeleton 1991, 20:69-78.
• [41]Finney M, Ruvkun G: The unc-86 gene product couples cell lineage and cell identity in C. elegans. Cell 1990, 63:895-905.