{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T13:15:12Z","timestamp":1753881312068,"version":"3.41.2"},"reference-count":31,"publisher":"American Society for Microbiology","issue":"5","license":[{"start":{"date-parts":[[2025,5,6]],"date-time":"2025-05-06T00:00:00Z","timestamp":1746489600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2025,5,6]],"date-time":"2025-05-06T00:00:00Z","timestamp":1746489600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["19H04326, 22H03802"],"award-info":[{"award-number":["19H04326, 22H03802"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["(19J21821"],"award-info":[{"award-number":["(19J21821"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100020959","name":"JST-Mirai Program","doi-asserted-by":"publisher","award":["JPMJMI22E8"],"award-info":[{"award-number":["JPMJMI22E8"]}],"id":[{"id":"10.13039\/501100020959","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Institution of Fermentation, Osaka","award":["G-2019-2-087"],"award-info":[{"award-number":["G-2019-2-087"]}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Microbiol Spectr"],"published-print":{"date-parts":[[2025,5,6]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:sec>\n            <jats:title\/>\n            <jats:p>\n              Methylotrophic yeasts can grow on methanol as the sole carbon and energy source. They have been used as hosts for heterologous protein production, which is achieved by their strongly and tightly regulated methanol-induced gene promoters. The expression levels of methanol-induced genes, such as\n              <jats:italic>AOD1<\/jats:italic>\n              and\n              <jats:italic>DAS1<\/jats:italic>\n              , are regulated by concentration-regulated methanol induction (CRMI). In this study, we identified CbMpp1 as a methanol-induced transcription factor necessary for the expression of multiple genes in the methylotrophic yeast\n              <jats:italic>Candida boidinii<\/jats:italic>\n              and identified the upstream activating sequences in the\n              <jats:italic>CbMPP1<\/jats:italic>\n              promoter. The expression of\n              <jats:italic>CbMPP1<\/jats:italic>\n              was induced by methanol under the control of CRMI and was dependent on other transcription factors, CbTrm1, CbTrm2, and CbHap complex but not on CbMpp1 itself. To investigate the physiological significance of the CRMI-dependent expression of\n              <jats:italic>CbMPP1<\/jats:italic>\n              , we constructed strains overexpressing or constitutively expressing\n              <jats:italic>CbMPP1<\/jats:italic>\n              with varying copy numbers. Analysis of growth with methanol and transcript levels of methanol-induced genes of these strains revealed that an appropriate amount of CbMpp1 was required for optimal growth with methanol and that excessive amounts of CbMpp1 repressed the expression of methanol-inducible genes. Such abnormal expression of\n              <jats:italic>CbMPP1<\/jats:italic>\n              in these strains caused accumulation of formaldehyde and growth defects on methanol. In addition, the\n              <jats:italic>Komagataella phaffii<\/jats:italic>\n              strain constitutively expressing\n              <jats:italic>KpMIT1<\/jats:italic>\n              (a homolog of\n              <jats:italic>CbMPP1<\/jats:italic>\n              ) at low levels exhibited retarded growth with methanol. Our results demonstrate that CbMpp1 and KpMit1, whose expression is regulated by CRMI, regulate the coordinated expression of multiple methanol-induced genes to achieve a balanced one-carbon metabolism.\n            <\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>IMPORTANCE<\/jats:title>\n            <jats:p>\n              Elucidating the molecular mechanism of how methanol-induced gene expression is regulated to attain a balanced one-carbon (C1) metabolism is important for determining the efficiency of heterologous protein production by methylotrophic yeasts. Although transcription factors involved in methanol-induced gene expression in these yeasts have been studied extensively, how these transcription factors regulate the expression of multiple methanol-induced genes in a concentration-regulated methanol induction (CRMI)-dependent manner has been unclear. In this study, we focused on the physiological significance of methanol inducibility of\n              <jats:italic>CbMpp1<\/jats:italic>\n              and\n              <jats:italic>KpMit1<\/jats:italic>\n              and found that CRMI-dependent\n              <jats:italic>CbMPP1<\/jats:italic>\n              and\n              <jats:italic>KpMIT1<\/jats:italic>\n              expressions are important for the coordinated regulation of multiple methanol-induced genes to facilitate a balanced C1 metabolism in the methylotrophic yeast.\n            <\/jats:p>\n          <\/jats:sec>","DOI":"10.1128\/spectrum.02853-24","type":"journal-article","created":{"date-parts":[[2025,3,31]],"date-time":"2025-03-31T13:20:02Z","timestamp":1743427202000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Methanol-induced transcription factor Mpp1 regulates the coordinated expression of multiple genes to achieve a balanced C1 metabolism in the methylotrophic yeast\n            <i>Candida boidinii<\/i>"],"prefix":"10.1128","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-2652-5052","authenticated-orcid":true,"given":"Koichi","family":"Inoue","sequence":"first","affiliation":[{"name":"Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-Oiwake, Sakyo-ku","place":["Kyoto, Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nono","family":"Saso","sequence":"additional","affiliation":[{"name":"Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-Oiwake, Sakyo-ku","place":["Kyoto, Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kosuke","family":"Iwase","sequence":"additional","affiliation":[{"name":"Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-Oiwake, Sakyo-ku","place":["Kyoto, Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhenyu","family":"Zhai","sequence":"additional","affiliation":[{"name":"Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-Oiwake, Sakyo-ku","place":["Kyoto, Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takahiro","family":"Tsuji","sequence":"additional","affiliation":[{"name":"Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-Oiwake, Sakyo-ku","place":["Kyoto, Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kazuyoshi","family":"Tabata","sequence":"additional","affiliation":[{"name":"Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-Oiwake, Sakyo-ku","place":["Kyoto, Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rie","family":"Sano","sequence":"additional","affiliation":[{"name":"Division of 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