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Its receptors, MTNR1A and MTNR1B, are members of the G\u2010protein\u2010coupled receptor (GPCR) family and are involved in sleep, circadian rhythm, and mood disorders, and in the inhibition of cancer growth. These receptors, therefore, represent significant molecular targets for insomnia, circadian sleep disorders, and cancer. The yeast <jats:italic>Saccharomyces cerevisiae<\/jats:italic> is an attractive host for assaying agonistic activity for human GPCR. We previously constructed a GPCR\u2010based biosensor employing a high\u2010sensitivity yeast strain that incorporated both a chimeric yeast\u2010human G\u03b1 protein and a bright fluorescent reporter gene (<jats:italic>ZsGreen<\/jats:italic>). Similar approaches have been used for simple and convenient measurements of various GPCR activities. In the current study, we constructed a fluorescence\u2010based yeast biosensor for monitoring the signaling activation of human melatonin receptors. We used this system to analyze point mutations, including previously unreported mutations of the consensus sequences of MTNR1A and MTNR1B melatonin receptors and compared their effects. Most mutations in the consensus sequences significantly affected the signaling capacities of both receptors, but several mutations showed differences between these subtype receptors. Thus, this yeast biosensor holds promise for revealing the functions of melatonin receptors.<\/jats:p>","DOI":"10.1002\/bit.27609","type":"journal-article","created":{"date-parts":[[2020,10,23]],"date-time":"2020-10-23T12:58:44Z","timestamp":1603457924000},"page":"863-876","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Comparative analyses of site\u2010directed mutagenesis of human melatonin MTNR1A and MTNR1B receptors using a yeast fluorescent biosensor"],"prefix":"10.1002","volume":"118","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0093-3754","authenticated-orcid":false,"given":"Yasuyuki","family":"Nakamura","sequence":"first","affiliation":[{"name":"Engineering Biology Research Center Kobe University Kobe Japan"},{"name":"Graduate School of Science, Technology and Innovation Kobe University Kobe Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ririka","family":"Asama","sequence":"additional","affiliation":[{"name":"Graduate School of Science, Technology and Innovation Kobe University Kobe Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takuya","family":"Tabata","sequence":"additional","affiliation":[{"name":"Graduate School of Science, Technology and Innovation Kobe University Kobe Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2156-4812","authenticated-orcid":false,"given":"Kenta","family":"Morita","sequence":"additional","affiliation":[{"name":"Department of Chemical Science and Engineering, Graduate School of Engineering Kobe University Kobe Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2428-1911","authenticated-orcid":false,"given":"Tatsuo","family":"Maruyama","sequence":"additional","affiliation":[{"name":"Department of Chemical Science and Engineering, Graduate School of Engineering Kobe University Kobe Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1527-5288","authenticated-orcid":false,"given":"Akihiko","family":"Kondo","sequence":"additional","affiliation":[{"name":"Engineering Biology Research Center Kobe University Kobe Japan"},{"name":"Graduate School of Science, Technology and Innovation Kobe University Kobe Japan"},{"name":"Department of Chemical Science and Engineering, Graduate School of Engineering Kobe University Kobe Japan"},{"name":"Center for Sustainable Resource Science RIKEN Yokohama Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2568-515X","authenticated-orcid":false,"given":"Jun","family":"Ishii","sequence":"additional","affiliation":[{"name":"Engineering Biology Research Center Kobe University Kobe Japan"},{"name":"Graduate School of Science, Technology and Innovation Kobe University Kobe Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2020,11,6]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jad.2015.07.017"},{"key":"e_1_2_8_3_1","first-page":"61","article-title":"Melatonin MT1 and MT2 receptor expression in Parkinson's disease","volume":"16","author":"Adi N.","year":"2010","journal-title":"Medical Science Monitor"},{"key":"e_1_2_8_4_1","first-page":"4693","article-title":"Melatonin inhibition of cancer growth in vivo involves suppression of tumor fatty acid metabolism via melatonin receptor\u2010mediated signal transduction events","volume":"59","author":"Blask D. 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