Warasirin Sornlek, Nattida Suwanakitti, Chutima Sonthirod, Sithichoke Tangphatsornruang, Supawadee Ingsriswang, Weerawat Runguphan, Lily Eurwilaichtr, Sutipa Tanapongpipat, Verawat Champreda, Niran Roongsawang, Peter J. Schaap, Vitor A. P. Martins dos Santos
{"title":"鉴定与酿酒酵母天然分离物 BCC39850 高温发酵特性相关的基因。","authors":"Warasirin Sornlek, Nattida Suwanakitti, Chutima Sonthirod, Sithichoke Tangphatsornruang, Supawadee Ingsriswang, Weerawat Runguphan, Lily Eurwilaichtr, Sutipa Tanapongpipat, Verawat Champreda, Niran Roongsawang, Peter J. Schaap, Vitor A. P. Martins dos Santos","doi":"10.1007/s00203-024-04117-x","DOIUrl":null,"url":null,"abstract":"<div><p>The fermentative model yeast <i>Saccharomyces cerevisiae</i> has been extensively used to study the genetic basis of stress response and homeostasis. In this study, we performed quantitative trait loci (QTL) analysis of the high-temperature fermentation trait of the progeny from the mating of the <i>S. cerevisiae</i> natural isolate BCC39850 (haploid#17) and the laboratory strain CEN.PK2-1C. A single QTL on chromosome X was identified, encompassing six candidate genes (<i>GEA1</i>, <i>PTK2</i>, <i>NTA1</i>, <i>NPA3</i>, <i>IRT1</i>, and <i>IML1</i>). The functions of these candidates were tested by reverse genetic experiments. Deletion mutants of <i>PTK2</i>, <i>NTA1</i>, and <i>IML1</i> showed growth defects at 42 °C. The <i>PTK2</i> knock-out mutant also showed significantly reduced ethanol production and plasma membrane H<sup>+</sup> ATPase activity and increased sensitivity to acetic acid, ethanol, amphotericin B (AMB), and <i>β</i>-1,3-glucanase treatment. The CRISPR-Cas9 system was used to construct knock-in mutants by replacement of <i>PTK2</i>, <i>NTA1</i>, <i>IML1</i>, and <i>NPA3</i> genes with BCC39850 alleles. The <i>PTK2</i> and <i>NTA1</i> knock-in mutants showed increased growth and ethanol production titers at 42 °C. These findings suggest an important role for the <i>PTK2</i> serine/threonine protein kinase in regulating plasma membrane H<sup>+</sup> ATPase activity and the <i>NTA1</i> N-terminal amidase in protein degradation via the ubiquitin-proteasome system machinery, which affects tolerance to heat stress in <i>S. cerevisiae</i>.</p></div>","PeriodicalId":8279,"journal":{"name":"Archives of Microbiology","volume":"206 10","pages":""},"PeriodicalIF":2.3000,"publicationDate":"2024-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Identification of genes associated with the high-temperature fermentation trait in the Saccharomyces cerevisiae natural isolate BCC39850\",\"authors\":\"Warasirin Sornlek, Nattida Suwanakitti, Chutima Sonthirod, Sithichoke Tangphatsornruang, Supawadee Ingsriswang, Weerawat Runguphan, Lily Eurwilaichtr, Sutipa Tanapongpipat, Verawat Champreda, Niran Roongsawang, Peter J. Schaap, Vitor A. P. Martins dos Santos\",\"doi\":\"10.1007/s00203-024-04117-x\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>The fermentative model yeast <i>Saccharomyces cerevisiae</i> has been extensively used to study the genetic basis of stress response and homeostasis. In this study, we performed quantitative trait loci (QTL) analysis of the high-temperature fermentation trait of the progeny from the mating of the <i>S. cerevisiae</i> natural isolate BCC39850 (haploid#17) and the laboratory strain CEN.PK2-1C. A single QTL on chromosome X was identified, encompassing six candidate genes (<i>GEA1</i>, <i>PTK2</i>, <i>NTA1</i>, <i>NPA3</i>, <i>IRT1</i>, and <i>IML1</i>). The functions of these candidates were tested by reverse genetic experiments. Deletion mutants of <i>PTK2</i>, <i>NTA1</i>, and <i>IML1</i> showed growth defects at 42 °C. The <i>PTK2</i> knock-out mutant also showed significantly reduced ethanol production and plasma membrane H<sup>+</sup> ATPase activity and increased sensitivity to acetic acid, ethanol, amphotericin B (AMB), and <i>β</i>-1,3-glucanase treatment. The CRISPR-Cas9 system was used to construct knock-in mutants by replacement of <i>PTK2</i>, <i>NTA1</i>, <i>IML1</i>, and <i>NPA3</i> genes with BCC39850 alleles. The <i>PTK2</i> and <i>NTA1</i> knock-in mutants showed increased growth and ethanol production titers at 42 °C. These findings suggest an important role for the <i>PTK2</i> serine/threonine protein kinase in regulating plasma membrane H<sup>+</sup> ATPase activity and the <i>NTA1</i> N-terminal amidase in protein degradation via the ubiquitin-proteasome system machinery, which affects tolerance to heat stress in <i>S. cerevisiae</i>.</p></div>\",\"PeriodicalId\":8279,\"journal\":{\"name\":\"Archives of Microbiology\",\"volume\":\"206 10\",\"pages\":\"\"},\"PeriodicalIF\":2.3000,\"publicationDate\":\"2024-09-04\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Archives of Microbiology\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://link.springer.com/article/10.1007/s00203-024-04117-x\",\"RegionNum\":3,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Archives of Microbiology","FirstCategoryId":"99","ListUrlMain":"https://link.springer.com/article/10.1007/s00203-024-04117-x","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"MICROBIOLOGY","Score":null,"Total":0}
Identification of genes associated with the high-temperature fermentation trait in the Saccharomyces cerevisiae natural isolate BCC39850
The fermentative model yeast Saccharomyces cerevisiae has been extensively used to study the genetic basis of stress response and homeostasis. In this study, we performed quantitative trait loci (QTL) analysis of the high-temperature fermentation trait of the progeny from the mating of the S. cerevisiae natural isolate BCC39850 (haploid#17) and the laboratory strain CEN.PK2-1C. A single QTL on chromosome X was identified, encompassing six candidate genes (GEA1, PTK2, NTA1, NPA3, IRT1, and IML1). The functions of these candidates were tested by reverse genetic experiments. Deletion mutants of PTK2, NTA1, and IML1 showed growth defects at 42 °C. The PTK2 knock-out mutant also showed significantly reduced ethanol production and plasma membrane H+ ATPase activity and increased sensitivity to acetic acid, ethanol, amphotericin B (AMB), and β-1,3-glucanase treatment. The CRISPR-Cas9 system was used to construct knock-in mutants by replacement of PTK2, NTA1, IML1, and NPA3 genes with BCC39850 alleles. The PTK2 and NTA1 knock-in mutants showed increased growth and ethanol production titers at 42 °C. These findings suggest an important role for the PTK2 serine/threonine protein kinase in regulating plasma membrane H+ ATPase activity and the NTA1 N-terminal amidase in protein degradation via the ubiquitin-proteasome system machinery, which affects tolerance to heat stress in S. cerevisiae.
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