Genome AnnouncGenome AnnouncgagaGAGenome Announcements2169-8287American Society for Microbiology1752 N St., N.W., Washington, DC237885523707601genomeA00392-1310.1128/genomeA.00392-13ProkaryotesDraft Genome of a Type 4 Pilus Defective Myxococcus xanthus Strain, DZF1MüllerSusanneaWillettJonathan W.bBahrSarah M.aScottJodie C.cWilsonJanet M.dDarnellCynthia L.aVlamakisHera C.eKirbyJohn R.aDepartment of Microbiology, University of Iowa, Iowa City, Iowa, USAUniversity of Chicago, Department of Biochemistry and Molecular Biology, Chicago, Ilinois, USAThe Forsyth Institute, Department of Microbiology, Cambridge, Massachusetts, USADivision of Select Agents and Toxins, Office of Public Health Preparedness and Response, Centers for Disease Control and Prevention, Atlanta, Georgia, USAHarvard Medical School, Department of Microbiology and Immunobiology, Boston, Massachusetts, USAAddress correspondence to John R. Kirby, john-kirby@uiowa.edu.

S.M., J.W.W., and S.M.B. contributed equally to this work.

2062013May-Jun201313e00392-139520131352013Copyright © 2013 Müller et al.2013Müller et al.This is an open-access article distributed under the terms of the Creative Commons Attribution 3.0 Unported license.

Myxococcus xanthus is a member of the Myxococcales order within the deltaproteobacterial subdivision. Here, we report the whole-genome shotgun sequence of the type IV pilus (T4P) defective strain DZF1, which includes many genes found in strain DZ2 but absent from strain DK1622.

cover-dateMay/June 2013access-typefree
GENOME ANNOUNCEMENT

Myxococcus xanthus is a soil-dwelling deltaproteobacterium with a genome length of >9.2 Mb. The Myxococcales were described in the late 19th century for their capacity to produce macroscopic sporangioles or fruiting bodies (1). Organization into fruiting bodies requires extracellular signaling and coordination of two genetically distinct motility systems (229). Previously, the sequence of M. xanthus strain DK1622 was determined (NC_008095.1) (30). Recently, we sequenced strain DZ2 (31). These two laboratory strains are noted for behavioral differences (2, 32, 33), and DZ2 has a larger genome (31).

Mxanthus DZF1 is directly descended from the intermediate strain DK101, the progenitor for DK1622, and displays reduced capacity for type IV pilus (T4P)-mediated motility. An earlier study (34) mapped two point mutations in DK101 to pilQ, which encodes the T4P secretin (G741S/N762G), accounting for some phenotypic differences between DZ2 or DK1622 and those strains harboring the pilQ allele. Because the DZ2 genome contains additional genes relative to DK1622 and because differences in motility have been noted, we sequenced DZF1 to determine if the additional genes were more likely gained by DZ2 or lost from DK1622, relative to the progenitor.

Mxanthus DZF1 was sequenced at the University of Iowa DNA Core Facility using 454 GS-FLX titanium technology. Chromosomal DNA was prepared as described previously (31) and processed for sequencing following established protocols. The resulting sequence comprises 388,477 reads totaling 249 Mb, representing 27-fold coverage. The genome was assembled de novo into 75 contigs using Newbler software version 2.7. The resulting genome is approximately 9.28 Mb, similar to that for DZ2 (31), and is approximately 147 kb larger than the DK1622 genome. The RAST annotation server (35) predicts a total of 7,704 coding sequences (CDS) within the DZF1 genome.

The M. xanthus DZF1 sequence reveals a single nucleotide polymorphism (SNP) in the pilQ gene, producing a G741S substitution, but lacks the N762G substitution found in DK101 (34). The impact of these SNPs has not been systematically determined but affects the interpretation of several previous studies. Indeed, deletion of mazF (encoding RNA interferase as part of a toxin-antitoxin system) is synthetic with the pilQ allele in both DZF1 and DK101 to affect cell death (36, 37).

Current analysis is ongoing to determine the role of genes found in DZF1, but not in DK1622, encoding proteins predicted to function in transcription, translation, signal transduction, fatty acid modification, and protein transport. Homologs to these genes are found in DZ2 as well as other myxobacteria, including Myxococcus fulvus, Stigmatella aurantiaca, and Sorangium cellulosum. The presence of sequences unique to both DZF1 and DZ2, while absent from DK1622, has been verified by PCR. Thus, the differences between DK1622 and both DZ2 and DZF1 are attributable to a loss of DNA from the DK1622 genome, likely following UV mutagenesis of DK101, which led to excision of one large prophage (38, 39) and may have induced additional lesions. We are investigating several unique sequences found in DZF1 and DZ2 for their role in M. xanthus biology.

Nucleotide sequence accession number.

This whole-genome shotgun project has been deposited at DDBJ/EMBL/GenBank under the accession number AOBT00000000. The version described in this paper is the first version.

Citation Müller S, Willett JW, Bahr SM, Scott JC, Wilson JM, Darnell CL, Vlamakis HC, Kirby JR. 2013. Draft genome of a type 4 pilus defective Myxococcus xanthus strain, DZF1. Genome Announc. 1(3):e00392-13. doi:10.1128/genomeA.00392-13.

ACKNOWLEDGMENTS

Support for this work was provided by the University of Iowa and NSF MCB-1244021 to J.R.K. Additional support for J.W.W. was provided by NIH T32 GM077973.

We thank Tom Willett, Tom Bair, and Kevin Knudtson for helpful discussions and data analysis.

The content is the responsibility of the authors and does not represent the official views of NIH, NSF, or the University of Iowa.

REFERENCES ThaxterR 1892 On the Myxobacteriaceae, a new order of Schizomycetes Bot. Gaz. 17:389406 BerlemanJEKirbyJR 2007 Multicellular development in Myxococcus xanthus is stimulated by predator-prey interactions. J. Bacteriol. 189:5675568217513469 BlackhartBDZusmanDR 1985 “Frizzy” genes of Myxococcus xanthus are involved in control of frequency of reversal of gliding motility. Proc. Natl. Acad. Sci. U. S. A. 82:876787703936045 BulyhaISchmidtCLenzPJakovljevicVHöneAMaierBHoppertMSøgaard-AndersenL 2009 Regulation of the type IV pili molecular machine by dynamic localization of two motor proteins. Mol. Microbiol. 74:69170619775250 CaberoyNBWelchRDJakobsenJSSlaterSCGarzaAG 2003 Global mutational analysis of NtrC-like activators in Myxococcus xanthus: identifying activator mutants defective for motility and fruiting body development. J. Bacteriol. 185:6083609414526020 CamposJMZusmanDR 1975 Regulation of development in Myxococcus xanthus: effect of 3′:5′-cyclic AMP, ADP, and nutrition. Proc. Natl. Acad. Sci. U. S. A. 72:518522164657 GiglioKMCaberoyNSuenGKaiserDGarzaAG 2011 A cascade of coregulating enhancer binding proteins initiates and propagates a multicellular developmental program. Proc. Natl. Acad. Sci. U. S. A. 108:E431E43921670274 HuntleySHamannNWegener-FeldbrüggeSTreuner-LangeAKubeMReinhardtRKlagesSMüllerRRonningCMNiermanWCSøgaard-AndersenL 2011 Comparative genomic analysis of fruiting body formation in Myxococcales. Mol. Biol. Evol. 28:1083109721037205 JelsbakLSøgaard-AndersenL 2002 Pattern formation by a cell surface-associated morphogen in Myxococcus xanthus. Proc. Natl. Acad. Sci. U. S. A. 99:2032203711842199 KirbyJRZusmanDR 2003 Chemosensory regulation of developmental gene expression in Myxococcus xanthus. Proc. Natl. Acad. Sci. U. S. A. 100:2008201312566562 KroosLHartzellPStephensKKaiserD 1988 A link between cell movement and gene expression argues that motility is required for cell-cell signaling during fruiting body development. Genes Dev. 2:167716853145903 KruseTLobedanzSBerthelsenNMSøgaard-AndersenL 2001 C-signal: a cell surface-associated morphogen that induces and co-ordinates multicellular fruiting body morphogenesis and sporulation in Myxococcus xanthus. Mol. Microbiol. 40:15616811298283 LucianoJAgrebiRLe GallAVWartelMFiegnaFDucretABrochier-ArmanetCMignotT 2011 Emergence and modular evolution of a novel motility machinery in bacteria. PLoS Genet. 7 e100226821931562 MacNeilSDMouzeyanAHartzellPL 1994 Genes required for both gliding motility and development in Myxococcus xanthus. Mol. Microbiol. 14:7857957891564 MaurielloEMMignotTYangZZusmanDR 2010 Gliding motility revisited: how do the myxobacteria move without flagella? Microbiol. Mol. Biol. Rev. 74:22924920508248 MignotT 2007 The elusive engine in Myxococcus xanthus gliding motility. Cell. Mol. Life Sci. 64:2733274517653507 MignotTKirbyJR 2008 Genetic circuitry controlling motility behaviors of Myxococcus xanthus. BioEssays 30:73374318623059 MignotTMerlieJPJrZusmanDR 2005 Regulated pole-to-pole oscillations of a bacterial gliding motility protein. Science 310:85585716272122 MignotTShaevitzJWHartzellPLZusmanDR 2007 Evidence that focal adhesion complexes power bacterial gliding motility. Science 315:85385617289998 NudlemanEWallDKaiserD 2005 Cell-to-cell transfer of bacterial outer membrane lipoproteins. Science 309:12512715994555 PathakDTWallD 2012 Identification of the cglC, cglD, cglE, and cglF genes and their role in cell contact-dependent gliding motility in Myxococcus xanthus. J. Bacteriol. 194:1940194922343295 ShimketsLJGillREKaiserD 1983 Developmental cell interactions in Myxococcus xanthus and the spoC locus. Proc. Natl. Acad. Sci. U. S. A. 80:1406141016593290 SingerMKaiserD 1995 Ectopic production of guanosine penta- and tetraphosphate can initiate early developmental gene expression in Myxococcus xanthus. Genes Dev. 9:163316447628697 WeiXPathakDTWallD 2011 Heterologous protein transfer within structured myxobacteria biofilms. Mol. Microbiol. 81:31532621635581 WelchRKaiserD 2001 Cell behavior in traveling wave patterns of myxobacteria. Proc. Natl. Acad. Sci. U. S. A. 98:149071491211752438 YouderianPBurkeNWhiteDJHartzellPL 2003 Identification of genes required for adventurous gliding motility in Myxococcus xanthus with the transposable element mariner. Mol. Microbiol. 49:55557012828649 YouderianPHartzellPL 2006 Transposon insertions of Magellan-4 that impair social gliding motility in Myxococcus xanthus. Genetics 172:1397141016299386 ZhangYDucretAShaevitzJMignotT 2012 From individual cell motility to collective behaviors: insights from a prokaryote, Myxococcus xanthus. FEMS Microbiol. Rev. 36:14916422091711 ZusmanDRScottAEYangZKirbyJR 2007 Chemosensory pathways, motility and development in Myxococcus xanthus. Nat. Rev. Microbiol. 5:86287217922045 GoldmanBSNiermanWCKaiserDSlaterSCDurkinASEisenJARonningCMBarbazukWBBlanchardMFieldCHallingCHinkleGIartchukOKimHSMackenzieCMadupuRMillerNShvartsbeynASullivanSAVaudinMWiegandRKaplanHBKaplanHB 2006 Evolution of sensory complexity recorded in a myxobacterial genome. Proc. Natl. Acad. Sci. U. S. A. 103:152001520517015832 MüllerSWillettJWBahrSMDarnellCLHummelsKRDongCKVlamakisHCKirbyJR 2013 Draft genome sequence of Myxococcus xanthus wild-type strain DZ2, a model organism for predation and development. Genome Announc. 1(3):e00217-13.10.1128/genomeA.00217-13 BerlemanJEChumleyTCheungPKirbyJR 2006 Rippling is a predatory behavior in Myxococcus xanthus. J. Bacteriol. 188:5888589516885457 O’ConnorKAZusmanDR 1988 Reexamination of the role of autolysis in the development of Myxococcus xanthus. J. Bacteriol. 170:410341123137213 WallDKolenbranderPEKaiserD 1999 The Myxococcus xanthus pilQ (sglA) gene encodes a secretin homolog required for type IV pilus biogenesis, social motility, and development. J. Bacteriol. 181:24339864308 AzizRKBartelsDBestAADeJonghMDiszTEdwardsRAFormsmaKGerdesSGlassEMKubalMMeyerFOlsenGJOlsonROstermanALOverbeekRAMcNeilLKPaarmannDPaczianTParrelloBPuschGDReichCStevensRVassievaOVonsteinVWilkeAZagnitkoO 2008 The RAST server: rapid annotations using subsystems technology. BMC Genomics 9:7518261238 BoyntonTOMcMurryJLShimketsLJ 2013 Characterization of Myxococcus xanthus MazF and implications for a new point of regulation. Mol. Microbiol. 87:1267127623369184 LeeBHolkenbrinkCTreuner-LangeAHiggsPI 2012 Myxococcus xanthus developmental cell fate production: heterogeneous accumulation of developmental regulatory proteins and reexamination of the role of MazF in developmental lysis. J. Bacteriol. 194:3058306822493014 ChenHKeselerIMShimketsLJ 1990 Genome size of Myxococcus xanthus determined by pulsed-field gel electrophoresis. J. Bacteriol. 172:420642132165472 ChenHWKuspaAKeselerIMShimketsLJ 1991 Physical map of the Myxococcus xanthus chromosome. J. Bacteriol. 173:210921151848221