Increased transcription of NOP15, involved in ribosome biogenesis in Saccharomyces cerevisiae, enhances the production yield of RNA as a source of nucleotide seasoning.

Yeast RNA is a good source of nucleotide seasoning, and more than half of yeast RNA consists of ribosomal RNA (rRNA). Previously, we reported the development of a Saccharomyces cerevisiae strain displaying a 1.4- to 2.3-times higher RNA content than the wild-type strain through the isolation of dominant suppressors (designated SupA to SupG strains) from a Δrrn10 disruptant showing decreased rRNA transcription. In the present study, the cloning of one of the genes responsible for the suppression was attempted using a genomic library from the SupD strain. NOP15, a gene involved in ribosome biogenesis, was found to be responsible for suppressing the growth defect of the Δrrn10 disruptant. The isolated NOP15 allele (designated NOP15(T-279C)) possessed a single T to C substitution at nucleotide position-279 of NOP15. The transcription level of NOP15(T-279C) in the originally isolated SupD strain was 2-fold higher than that in the Δrrn10 disruptant. Furthermore, a dose-dependent relationship between the transcription level of NOP15 and total amount of RNA in the Δrrn10 disruptant was observed: the enhanced transcription due to the NOP15(T-279C) allele is involved in the suppression mechanisms in the SupD strain. Introduction of the NOP15(T-279C) allele into the wild-type strain increased the total RNA content by 1.4-fold. These results indicate that the transcription level of NOP15 is an important determinant of the productivity of RNA and that its increased transcription provides an effective approach to obtain higher RNA yields in yeast.

[1]  T. Petes Yeast ribosomal DNA genes are located on chromosome XII. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[2]  R. Sikorski,et al.  A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. , 1989, Genetics.

[3]  T. Nagodawithana Yeast-derived flavors and flavor enhancers and their probable mode of action , 1992 .

[4]  R. Courbeyrette,et al.  Yaf9, a Novel NuA4 Histone Acetyltransferase Subunit, Is Required for the Cellular Response to Spindle Stress in Yeast , 2003, Molecular and Cellular Biology.

[5]  Anuj Kumar,et al.  Large-scale analysis of yeast filamentous growth by systematic gene disruption and overexpression. , 2008, Molecular biology of the cell.

[6]  J. Shabanowitz,et al.  Composition and functional characterization of yeast 66S ribosome assembly intermediates. , 2001, Molecular cell.

[7]  M. Nomura,et al.  Transcription of Multiple Yeast Ribosomal DNA Genes Requires Targeting of UAF to the Promoter by Uaf30 , 2008, Molecular and Cellular Biology.

[8]  S. M. Sullivan,et al.  In vivo functional characterization of the Saccharomyces cerevisiae 60S biogenesis GTPase Nog1 , 2007, Molecular Genetics and Genomics.

[9]  Y. Nogi,et al.  Transcription of rDNA in the Yeast Saccharomyces cerevisiae , 2013 .

[10]  M. Nomura,et al.  Interaction of TATA-Binding Protein with Upstream Activation Factor Is Required for Activated Transcription of Ribosomal DNA by RNA Polymerase I in Saccharomyces cerevisiae In Vivo , 1998, Molecular and Cellular Biology.

[11]  J. Warner,et al.  The economics of ribosome biosynthesis in yeast. , 1999, Trends in biochemical sciences.

[12]  M. Nomura Ribosomal RNA genes, RNA polymerases, nucleolar structures, and synthesis of rRNA in the yeast Saccharomyces cerevisiae. , 2001, Cold Spring Harbor symposia on quantitative biology.

[13]  Y. Nogi,et al.  Multiprotein transcription factor UAF interacts with the upstream element of the yeast RNA polymerase I promoter and forms a stable preinitiation complex. , 1996, Genes & development.

[14]  Y. Kaneko,et al.  Chromosome XII context is important for rDNA function in yeast , 2006, Nucleic Acids Research.

[15]  A. López-Munguía,et al.  Continuous production of 5′-ribonucleotides from yeast RNA by hydrolysis with immobilized 5′-phosphodiesterase and 5′-adenylate deaminase , 1994, World journal of microbiology & biotechnology.

[16]  P. Gleizes,et al.  Processing of 20S pre‐rRNA to 18S ribosomal RNA in yeast requires Rrp10p, an essential non‐ribosomal cytoplasmic protein , 2001, The EMBO journal.

[17]  Y. Kaneko,et al.  Construction of a Saccharomyces cerevisiae strain with a high level of RNA. , 2011, Journal of bioscience and bioengineering.

[18]  Y. Nogi,et al.  RRN3 gene of Saccharomyces cerevisiae encodes an essential RNA polymerase I transcription factor which interacts with the polymerase independently of DNA template. , 1996, The EMBO journal.

[19]  S. Campanaro,et al.  Gene disruption and basic phenotypic analysis of nine novel yeast genes from chromosome XIV , 2000, Yeast.

[20]  J. Strathern,et al.  Methods in yeast genetics : a Cold Spring Harbor Laboratory course manual , 2005 .

[21]  L. Lindahl,et al.  The RNA of RNase MRP is required for normal processing of ribosomal RNA. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[22]  David Tollervey,et al.  Yeast Nop15p is an RNA‐binding protein required for pre‐rRNA processing and cytokinesis , 2003, The EMBO journal.