Genetic analysis of a type IV pili-like locus in the archaeon Methanococcus maripaludis

• Published in: Archives of microbiology Vol. 196; no. 3; pp. 179 - 191
• Main Authors: Nair, Divya B, Uchida, Kaoru, Aizawa, Shin-Ichi, Jarrell, Ken F
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer-Verlag 01.03.2014; Springer Berlin Heidelberg; Springer Nature B.V
• Subjects: Adenosine Triphosphatases - genetics; Adenosine Triphosphatases - metabolism; adenosinetriphosphatase; Biochemistry; Biomedical and Life Sciences; Biotechnology; Cell Biology; Cell Membrane - genetics; Conserved Sequence - genetics; Ecology; electron microscopy; fimbriae; Fimbriae Proteins - genetics; Fimbriae Proteins - metabolism; Gene Deletion; Genes; Genetic Complementation Test; Genomes; Life Sciences; loci; Methanococcus - enzymology; Methanococcus - genetics; Methanococcus - ultrastructure; Methanococcus maripaludis; Microbial Ecology; Microbiology; multigene family; Operon - genetics; Original Paper; Peptides; Plasmids - genetics; Proteins; Reverse Transcriptase Polymerase Chain Reaction; transcription (genetics); Prepilin peptidase; Type IV pili; ATPase; Archaea
• ISSN: 0302-8933; 1432-072X
• DOI: 10.1007/s00203-014-0956-4

Methanococcus maripaludis is a stringently anaerobic archaeon with two studied surface structures, archaella and type IV pili. Previously, it was shown that three pilin genes (mmp0233 [epdA], mmp0236 [epdB] and mmp0237 [epdC]) located within an 11 gene cluster in the genome were necessary for normal piliation. This study focused on analysis of the remaining genes to determine their potential involvement in piliation. Reverse transcriptase PCR experiments demonstrated the 11 genes formed a single transcriptional unit. Deletions were made in all the non-pilin genes except mmp0231. Electron microscopy revealed that all the genes in the locus except mmp0235 and mmp0238 were essential for piliation. Complementation with a plasmid-borne wild-type copy of the deleted gene restored at least some piliation. We identified genes for an assembly ATPase and two versions of the conserved pilin platform forming protein necessary for pili assembly at a separate genetic locus.