Mutational analysis of action of mitochondrial fusion factor mitofusin-2
Mitofusin-2 (Mfn2) is an essential component of mitochondrial fusion machinery, but its molecular mechanism of action is not clear. We found that a Mfn2 deletion mutant lacking two transmembrane spans (Mfn[superscript [Delta]TM]) acts as a dominant-negative mutant and blocks mitochondrial fusion. Fu...
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| Veröffentlicht in: | Journal of cell science 2005-07, Vol.118 (14), p.3153-3161 |
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| container_title | Journal of cell science |
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| creator | Honda, Shinji Aihara, Takeshi Hontani, Masayasu Okubo, Katsuhiko Hirose, Shigehisa |
| description | Mitofusin-2 (Mfn2) is an essential component of mitochondrial fusion machinery, but its molecular mechanism of action is not clear. We found that a Mfn2 deletion mutant lacking two transmembrane spans (Mfn[superscript [Delta]TM]) acts as a dominant-negative mutant and blocks mitochondrial fusion. Furthermore, detailed analysis of various mutants of Mfn[superscript [Delta]TM] revealed that GTPase activity and four regions highly conserved from nematodes to mammals are necessary for the dominant-negative effect. Immunoprecipitation studies of the N- and C-terminal cytosolic tails of Mfn2 showed that in addition to the coiled-coil domains previously identified, a highly conserved domain in the most N-terminal region and GTPase activity are necessary for the interaction between the N- and C-terminal tails, which is in turn required for the dominant-negative effect. In addition, we found unexpectedly that overexpression of the deletion mutant composed of one short region each in the N- and C-terminal tails of Mfn2 resulted in loss of mitochondrial membrane potential, suggesting that Mfn2 might also be connected to maintenance of mitochondrial membrane potential. |
| doi_str_mv | 10.1242/jcs.02449 |
| format | Article |
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We found that a Mfn2 deletion mutant lacking two transmembrane spans (Mfn[superscript [Delta]TM]) acts as a dominant-negative mutant and blocks mitochondrial fusion. Furthermore, detailed analysis of various mutants of Mfn[superscript [Delta]TM] revealed that GTPase activity and four regions highly conserved from nematodes to mammals are necessary for the dominant-negative effect. Immunoprecipitation studies of the N- and C-terminal cytosolic tails of Mfn2 showed that in addition to the coiled-coil domains previously identified, a highly conserved domain in the most N-terminal region and GTPase activity are necessary for the interaction between the N- and C-terminal tails, which is in turn required for the dominant-negative effect. 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We found that a Mfn2 deletion mutant lacking two transmembrane spans (Mfn[superscript [Delta]TM]) acts as a dominant-negative mutant and blocks mitochondrial fusion. Furthermore, detailed analysis of various mutants of Mfn[superscript [Delta]TM] revealed that GTPase activity and four regions highly conserved from nematodes to mammals are necessary for the dominant-negative effect. Immunoprecipitation studies of the N- and C-terminal cytosolic tails of Mfn2 showed that in addition to the coiled-coil domains previously identified, a highly conserved domain in the most N-terminal region and GTPase activity are necessary for the interaction between the N- and C-terminal tails, which is in turn required for the dominant-negative effect. In addition, we found unexpectedly that overexpression of the deletion mutant composed of one short region each in the N- and C-terminal tails of Mfn2 resulted in loss of mitochondrial membrane potential, suggesting that Mfn2 might also be connected to maintenance of mitochondrial membrane potential.</description><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Cell Line</subject><subject>Cercopithecus aethiops</subject><subject>Conserved Sequence</subject><subject>COS Cells</subject><subject>Gene Deletion</subject><subject>GTP Phosphohydrolases - genetics</subject><subject>GTP Phosphohydrolases - metabolism</subject><subject>Humans</subject><subject>Immunoprecipitation</subject><subject>Intracellular Membranes - enzymology</subject><subject>Intracellular Membranes - physiology</subject><subject>Mammalia</subject><subject>Membrane Potentials - physiology</subject><subject>Mice</subject><subject>Mitochondria - enzymology</subject><subject>Mitochondria - genetics</subject><subject>Mitochondria - physiology</subject><subject>Molecular Sequence Data</subject><subject>Mutation</subject><subject>Nematoda</subject><subject>Protein Structure, Tertiary</subject><subject>Transfection</subject><issn>0021-9533</issn><issn>1477-9137</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2005</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkLFOwzAQhi0EoqUw8ALQCYkh5c524nhEFVCkIgbobDmODamSutjJ0LcnaSsxstyd_v_TDR8h1wgzpJw-rE2cAeVcnpAxciESiUyckjEAxUSmjI3IRYxrABBUinMywlTmKc_YmCzeula3ld_oeqr7sYtVnHo31WYIh6upWm--_aYMVc-4Lg6563sf9t2QbBJ6Sc6crqO9Ou4JWT0_fc4XyfL95XX-uEwMT3mbGJ1zpFAYcBplRnlRMMy5K6jQRamdSS1mKLk1wLISXVGYErhkZZnnhruUTcjd4e82-J_OxlY1VTS2rvXG-i6qLAeaIcN_QRRMZBQH8P4AmuBjDNapbagaHXYKQQ1-Ve9X7f327M3xaVc0tvwjj0J74PYAOO2V_gpVVKsPCsgAAZEjZ7_8BH-X</recordid><startdate>20050715</startdate><enddate>20050715</enddate><creator>Honda, Shinji</creator><creator>Aihara, Takeshi</creator><creator>Hontani, Masayasu</creator><creator>Okubo, Katsuhiko</creator><creator>Hirose, Shigehisa</creator><scope>FBQ</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>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>20050715</creationdate><title>Mutational analysis of action of mitochondrial fusion factor mitofusin-2</title><author>Honda, Shinji ; Aihara, Takeshi ; Hontani, Masayasu ; Okubo, Katsuhiko ; Hirose, Shigehisa</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c454t-ca84120bc0fa19624bb3184fb27abdafc5e16194ec036d1fbbcd0493dd88c4f53</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2005</creationdate><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Cell Line</topic><topic>Cercopithecus aethiops</topic><topic>Conserved Sequence</topic><topic>COS Cells</topic><topic>Gene Deletion</topic><topic>GTP Phosphohydrolases - genetics</topic><topic>GTP Phosphohydrolases - metabolism</topic><topic>Humans</topic><topic>Immunoprecipitation</topic><topic>Intracellular Membranes - enzymology</topic><topic>Intracellular Membranes - physiology</topic><topic>Mammalia</topic><topic>Membrane Potentials - physiology</topic><topic>Mice</topic><topic>Mitochondria - enzymology</topic><topic>Mitochondria - genetics</topic><topic>Mitochondria - physiology</topic><topic>Molecular Sequence Data</topic><topic>Mutation</topic><topic>Nematoda</topic><topic>Protein Structure, Tertiary</topic><topic>Transfection</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Honda, Shinji</creatorcontrib><creatorcontrib>Aihara, Takeshi</creatorcontrib><creatorcontrib>Hontani, Masayasu</creatorcontrib><creatorcontrib>Okubo, Katsuhiko</creatorcontrib><creatorcontrib>Hirose, Shigehisa</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of cell science</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Honda, Shinji</au><au>Aihara, Takeshi</au><au>Hontani, Masayasu</au><au>Okubo, Katsuhiko</au><au>Hirose, Shigehisa</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Mutational analysis of action of mitochondrial fusion factor mitofusin-2</atitle><jtitle>Journal of cell science</jtitle><addtitle>J Cell Sci</addtitle><date>2005-07-15</date><risdate>2005</risdate><volume>118</volume><issue>14</issue><spage>3153</spage><epage>3161</epage><pages>3153-3161</pages><issn>0021-9533</issn><eissn>1477-9137</eissn><abstract>Mitofusin-2 (Mfn2) is an essential component of mitochondrial fusion machinery, but its molecular mechanism of action is not clear. We found that a Mfn2 deletion mutant lacking two transmembrane spans (Mfn[superscript [Delta]TM]) acts as a dominant-negative mutant and blocks mitochondrial fusion. Furthermore, detailed analysis of various mutants of Mfn[superscript [Delta]TM] revealed that GTPase activity and four regions highly conserved from nematodes to mammals are necessary for the dominant-negative effect. Immunoprecipitation studies of the N- and C-terminal cytosolic tails of Mfn2 showed that in addition to the coiled-coil domains previously identified, a highly conserved domain in the most N-terminal region and GTPase activity are necessary for the interaction between the N- and C-terminal tails, which is in turn required for the dominant-negative effect. In addition, we found unexpectedly that overexpression of the deletion mutant composed of one short region each in the N- and C-terminal tails of Mfn2 resulted in loss of mitochondrial membrane potential, suggesting that Mfn2 might also be connected to maintenance of mitochondrial membrane potential.</abstract><cop>England</cop><pmid>15985463</pmid><doi>10.1242/jcs.02449</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Company of Biologists |
| subjects | Amino Acid Sequence Animals Cell Line Cercopithecus aethiops Conserved Sequence COS Cells Gene Deletion GTP Phosphohydrolases - genetics GTP Phosphohydrolases - metabolism Humans Immunoprecipitation Intracellular Membranes - enzymology Intracellular Membranes - physiology Mammalia Membrane Potentials - physiology Mice Mitochondria - enzymology Mitochondria - genetics Mitochondria - physiology Molecular Sequence Data Mutation Nematoda Protein Structure, Tertiary Transfection |
| title | Mutational analysis of action of mitochondrial fusion factor mitofusin-2 |
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