Novel Component of the Rhodobacter sphaeroides Fla1 Flagellum Is Essential for Motor Rotation
Here we describe a novel component essential for flagellar rotation in Rhodobacter sphaeroides. This protein is encoded by motF (RSP_0067), the first gene of a predicted transcriptional unit which contains two hypothetical genes. Sequence analysis indicated that MotF is a bitopic membrane-spanning p...
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| Veröffentlicht in: | Journal of Bacteriology 2012-11, Vol.194 (22), p.6174-6183 |
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| creator | Ramírez-Cabrera, Victor Poggio, Sebastian Domenzain, Clelia Osorio, Aurora Dreyfus, Georges Camarena, Laura |
| description | Here we describe a novel component essential for flagellar rotation in Rhodobacter sphaeroides. This protein is encoded by motF (RSP_0067), the first gene of a predicted transcriptional unit which contains two hypothetical genes. Sequence analysis indicated that MotF is a bitopic membrane-spanning protein. Protease sensitivity assays and green fluorescent protein (GFP) fusions confirmed this prediction and allowed us to conclude that the C terminus of MotF is located in the periplasmic space. Wild-type cells expressing a functional GFP-MotF fusion show a single fluorescent focus per cell. The localization of this protein in different genetic backgrounds allowed us to determine that normal localization of MotF depends on the presence of FliL and MotB. Characterization of a ΔmotF pseudorevertant strain revealed that a single nucleotide change in motB suppresses the Mot− phenotype of the motF mutant. Additionally, we show that MotF also becomes dispensable when other mutant alleles of motB previously isolated as second-site suppressors of ΔfliL were expressed in the motF mutant strain. These results show that MotF is a new component of the Fla1 flagellum, which together with FliL is required to promote flagellar rotation, possibly through MotB. |
| doi_str_mv | 10.1128/jb.00850-12 |
| format | Article |
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This protein is encoded by motF (RSP_0067), the first gene of a predicted transcriptional unit which contains two hypothetical genes. Sequence analysis indicated that MotF is a bitopic membrane-spanning protein. Protease sensitivity assays and green fluorescent protein (GFP) fusions confirmed this prediction and allowed us to conclude that the C terminus of MotF is located in the periplasmic space. Wild-type cells expressing a functional GFP-MotF fusion show a single fluorescent focus per cell. The localization of this protein in different genetic backgrounds allowed us to determine that normal localization of MotF depends on the presence of FliL and MotB. Characterization of a ΔmotF pseudorevertant strain revealed that a single nucleotide change in motB suppresses the Mot− phenotype of the motF mutant. Additionally, we show that MotF also becomes dispensable when other mutant alleles of motB previously isolated as second-site suppressors of ΔfliL were expressed in the motF mutant strain. These results show that MotF is a new component of the Fla1 flagellum, which together with FliL is required to promote flagellar rotation, possibly through MotB.</description><identifier>ISSN: 0021-9193</identifier><identifier>EISSN: 1098-5530</identifier><identifier>EISSN: 1067-8832</identifier><identifier>DOI: 10.1128/jb.00850-12</identifier><identifier>PMID: 22961858</identifier><identifier>CODEN: JOBAAY</identifier><language>eng</language><publisher>Washington, DC: American Society for Microbiology</publisher><subject>alleles ; Amino Acid Sequence ; Animals ; Antibodies, Bacterial ; Bacterial proteins ; Bacterial Proteins - genetics ; Bacterial Proteins - metabolism ; Bacteriology ; Biological and medical sciences ; Cells ; Female ; Flagella - genetics ; Flagella - metabolism ; flagellum ; fluorescence ; Fundamental and applied biological sciences. Psychology ; Gene expression ; Gene Expression Regulation, Bacterial - physiology ; genetic background ; Genotype & phenotype ; green fluorescent protein ; Membrane Proteins - genetics ; Membrane Proteins - metabolism ; Mice ; Mice, Inbred BALB C ; Microbiology ; Miscellaneous ; Molecular Motor Proteins - genetics ; Molecular Motor Proteins - metabolism ; Motor ability ; mutants ; Mutation ; Periplasm ; phenotype ; Plasmids ; Point Mutation ; prediction ; proteinases ; Rhodobacter sphaeroides ; Rhodobacter sphaeroides - genetics ; Rhodobacter sphaeroides - metabolism ; sequence analysis ; transcription (genetics)</subject><ispartof>Journal of Bacteriology, 2012-11, Vol.194 (22), p.6174-6183</ispartof><rights>2015 INIST-CNRS</rights><rights>Copyright American Society for Microbiology Nov 2012</rights><rights>Copyright © 2012, American Society for Microbiology. All Rights Reserved. 2012 American Society for Microbiology</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c589t-e6fae04bc5c8522263ce141a05976fc4b5491cad86c5eed42a6bd3c9df2933843</citedby><cites>FETCH-LOGICAL-c589t-e6fae04bc5c8522263ce141a05976fc4b5491cad86c5eed42a6bd3c9df2933843</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3486363/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3486363/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,723,776,780,881,27901,27902,53766,53768</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=26598741$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22961858$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ramírez-Cabrera, Victor</creatorcontrib><creatorcontrib>Poggio, Sebastian</creatorcontrib><creatorcontrib>Domenzain, Clelia</creatorcontrib><creatorcontrib>Osorio, Aurora</creatorcontrib><creatorcontrib>Dreyfus, Georges</creatorcontrib><creatorcontrib>Camarena, Laura</creatorcontrib><title>Novel Component of the Rhodobacter sphaeroides Fla1 Flagellum Is Essential for Motor Rotation</title><title>Journal of Bacteriology</title><addtitle>J Bacteriol</addtitle><description>Here we describe a novel component essential for flagellar rotation in Rhodobacter sphaeroides. This protein is encoded by motF (RSP_0067), the first gene of a predicted transcriptional unit which contains two hypothetical genes. Sequence analysis indicated that MotF is a bitopic membrane-spanning protein. Protease sensitivity assays and green fluorescent protein (GFP) fusions confirmed this prediction and allowed us to conclude that the C terminus of MotF is located in the periplasmic space. Wild-type cells expressing a functional GFP-MotF fusion show a single fluorescent focus per cell. The localization of this protein in different genetic backgrounds allowed us to determine that normal localization of MotF depends on the presence of FliL and MotB. Characterization of a ΔmotF pseudorevertant strain revealed that a single nucleotide change in motB suppresses the Mot− phenotype of the motF mutant. Additionally, we show that MotF also becomes dispensable when other mutant alleles of motB previously isolated as second-site suppressors of ΔfliL were expressed in the motF mutant strain. These results show that MotF is a new component of the Fla1 flagellum, which together with FliL is required to promote flagellar rotation, possibly through MotB.</description><subject>alleles</subject><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Antibodies, Bacterial</subject><subject>Bacterial proteins</subject><subject>Bacterial Proteins - genetics</subject><subject>Bacterial Proteins - metabolism</subject><subject>Bacteriology</subject><subject>Biological and medical sciences</subject><subject>Cells</subject><subject>Female</subject><subject>Flagella - genetics</subject><subject>Flagella - metabolism</subject><subject>flagellum</subject><subject>fluorescence</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene expression</subject><subject>Gene Expression Regulation, Bacterial - physiology</subject><subject>genetic background</subject><subject>Genotype & phenotype</subject><subject>green fluorescent protein</subject><subject>Membrane Proteins - genetics</subject><subject>Membrane Proteins - metabolism</subject><subject>Mice</subject><subject>Mice, Inbred BALB C</subject><subject>Microbiology</subject><subject>Miscellaneous</subject><subject>Molecular Motor Proteins - genetics</subject><subject>Molecular Motor Proteins - metabolism</subject><subject>Motor ability</subject><subject>mutants</subject><subject>Mutation</subject><subject>Periplasm</subject><subject>phenotype</subject><subject>Plasmids</subject><subject>Point Mutation</subject><subject>prediction</subject><subject>proteinases</subject><subject>Rhodobacter sphaeroides</subject><subject>Rhodobacter sphaeroides - genetics</subject><subject>Rhodobacter sphaeroides - metabolism</subject><subject>sequence analysis</subject><subject>transcription (genetics)</subject><issn>0021-9193</issn><issn>1098-5530</issn><issn>1067-8832</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqN0k1v1DAQBuAIgehSOHEHq6gSEkrx-GvtCxKsWigqIBV6RJbjOBuvknixk1b8exx2KR8nLvbBj1_NeFwUjwGfABD5clOdYCw5LoHcKRaAlSw5p_huscCYQKlA0YPiQUobjIExTu4XB4QoAZLLRfH1Y7h2HVqFfhsGN4woNGhsHbpsQx0qY0cXUdq2xsXga5fQWWdgXtau66YenSd0mlK-502HmhDRhzDm9TKMZvRheFjca0yX3KP9flhcnZ1-Wb0rLz69PV-9vigtl2osnWiMw6yy3EpOCBHUOmBgMFdL0VhWcabAmloKy52rGTGiqqlVdUMUpZLRw-LVLnc7Vb2rbS4omk5vo-9N_K6D8frvk8G3eh2uNWVSUEFzwPN9QAzfJpdG3ftkc49mcGFKGsQyVyYo-Q8KuQEiQMxlPfuHbsIUh_wSs2IME05VVi92ysaQUnTNbd2AZyf1-zf654Q1kKyf_Nnqrf010gyO98Aka7ommsH69NsJruSSQXZHO9f6dXvjo9Mm9XpTaVAsp2kBy7mDpzvUmKDNOuagq88EA5__kqJLoD8AKh3B0w</recordid><startdate>20121101</startdate><enddate>20121101</enddate><creator>Ramírez-Cabrera, Victor</creator><creator>Poggio, Sebastian</creator><creator>Domenzain, Clelia</creator><creator>Osorio, Aurora</creator><creator>Dreyfus, Georges</creator><creator>Camarena, Laura</creator><general>American Society for Microbiology</general><scope>FBQ</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7TM</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>7S9</scope><scope>L.6</scope><scope>5PM</scope></search><sort><creationdate>20121101</creationdate><title>Novel Component of the Rhodobacter sphaeroides Fla1 Flagellum Is Essential for Motor Rotation</title><author>Ramírez-Cabrera, Victor ; Poggio, Sebastian ; Domenzain, Clelia ; Osorio, Aurora ; Dreyfus, Georges ; Camarena, Laura</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c589t-e6fae04bc5c8522263ce141a05976fc4b5491cad86c5eed42a6bd3c9df2933843</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>alleles</topic><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Antibodies, Bacterial</topic><topic>Bacterial proteins</topic><topic>Bacterial Proteins - genetics</topic><topic>Bacterial Proteins - metabolism</topic><topic>Bacteriology</topic><topic>Biological and medical sciences</topic><topic>Cells</topic><topic>Female</topic><topic>Flagella - genetics</topic><topic>Flagella - metabolism</topic><topic>flagellum</topic><topic>fluorescence</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene expression</topic><topic>Gene Expression Regulation, Bacterial - physiology</topic><topic>genetic background</topic><topic>Genotype & phenotype</topic><topic>green fluorescent protein</topic><topic>Membrane Proteins - genetics</topic><topic>Membrane Proteins - metabolism</topic><topic>Mice</topic><topic>Mice, Inbred BALB C</topic><topic>Microbiology</topic><topic>Miscellaneous</topic><topic>Molecular Motor Proteins - genetics</topic><topic>Molecular Motor Proteins - metabolism</topic><topic>Motor ability</topic><topic>mutants</topic><topic>Mutation</topic><topic>Periplasm</topic><topic>phenotype</topic><topic>Plasmids</topic><topic>Point Mutation</topic><topic>prediction</topic><topic>proteinases</topic><topic>Rhodobacter sphaeroides</topic><topic>Rhodobacter sphaeroides - genetics</topic><topic>Rhodobacter sphaeroides - metabolism</topic><topic>sequence analysis</topic><topic>transcription (genetics)</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ramírez-Cabrera, Victor</creatorcontrib><creatorcontrib>Poggio, Sebastian</creatorcontrib><creatorcontrib>Domenzain, Clelia</creatorcontrib><creatorcontrib>Osorio, Aurora</creatorcontrib><creatorcontrib>Dreyfus, Georges</creatorcontrib><creatorcontrib>Camarena, Laura</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>AGRICOLA</collection><collection>AGRICOLA - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Journal of Bacteriology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ramírez-Cabrera, Victor</au><au>Poggio, Sebastian</au><au>Domenzain, Clelia</au><au>Osorio, Aurora</au><au>Dreyfus, Georges</au><au>Camarena, Laura</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Novel Component of the Rhodobacter sphaeroides Fla1 Flagellum Is Essential for Motor Rotation</atitle><jtitle>Journal of Bacteriology</jtitle><addtitle>J Bacteriol</addtitle><date>2012-11-01</date><risdate>2012</risdate><volume>194</volume><issue>22</issue><spage>6174</spage><epage>6183</epage><pages>6174-6183</pages><issn>0021-9193</issn><eissn>1098-5530</eissn><eissn>1067-8832</eissn><coden>JOBAAY</coden><abstract>Here we describe a novel component essential for flagellar rotation in Rhodobacter sphaeroides. This protein is encoded by motF (RSP_0067), the first gene of a predicted transcriptional unit which contains two hypothetical genes. Sequence analysis indicated that MotF is a bitopic membrane-spanning protein. Protease sensitivity assays and green fluorescent protein (GFP) fusions confirmed this prediction and allowed us to conclude that the C terminus of MotF is located in the periplasmic space. Wild-type cells expressing a functional GFP-MotF fusion show a single fluorescent focus per cell. The localization of this protein in different genetic backgrounds allowed us to determine that normal localization of MotF depends on the presence of FliL and MotB. Characterization of a ΔmotF pseudorevertant strain revealed that a single nucleotide change in motB suppresses the Mot− phenotype of the motF mutant. Additionally, we show that MotF also becomes dispensable when other mutant alleles of motB previously isolated as second-site suppressors of ΔfliL were expressed in the motF mutant strain. These results show that MotF is a new component of the Fla1 flagellum, which together with FliL is required to promote flagellar rotation, possibly through MotB.</abstract><cop>Washington, DC</cop><pub>American Society for Microbiology</pub><pmid>22961858</pmid><doi>10.1128/jb.00850-12</doi><tpages>10</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | alleles Amino Acid Sequence Animals Antibodies, Bacterial Bacterial proteins Bacterial Proteins - genetics Bacterial Proteins - metabolism Bacteriology Biological and medical sciences Cells Female Flagella - genetics Flagella - metabolism flagellum fluorescence Fundamental and applied biological sciences. Psychology Gene expression Gene Expression Regulation, Bacterial - physiology genetic background Genotype & phenotype green fluorescent protein Membrane Proteins - genetics Membrane Proteins - metabolism Mice Mice, Inbred BALB C Microbiology Miscellaneous Molecular Motor Proteins - genetics Molecular Motor Proteins - metabolism Motor ability mutants Mutation Periplasm phenotype Plasmids Point Mutation prediction proteinases Rhodobacter sphaeroides Rhodobacter sphaeroides - genetics Rhodobacter sphaeroides - metabolism sequence analysis transcription (genetics) |
| title | Novel Component of the Rhodobacter sphaeroides Fla1 Flagellum Is Essential for Motor Rotation |
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