Ribosomal protein L30 is dispensable in the yeast Saccharomyces cerevisiae

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Published inMolecular and Cellular Biology Vol. 10; no. 10; pp. 5235 - 5243
Main Authors Baronas-Lowell, D M, Warner, J R
Format Journal Article
LanguageEnglish
Published Washington, DC American Society for Microbiology 01.10.1990
Subjects
Amino Acid Sequence
Base Sequence
Biological and medical sciences
Blotting, Northern
Cloning, Molecular
DNA Mutational Analysis
Electrophoresis, Gel, Two-Dimensional
Fundamental and applied biological sciences. Psychology
Genes, Fungal
Microbiology
Molecular Sequence Data
Mycology
Polymerase Chain Reaction
Polyribosomes - metabolism
Restriction Mapping
Ribosomal Proteins - physiology
Ribosomes - physiology
Ribosomes - ultrastructure
RNA Splicing
RNA, Viral - genetics
Saccharomyces cerevisiae
Saccharomyces cerevisiae - physiology
Structure-Activity Relationship
Fungi
Gene
Nucleotide sequence
Ascomycetes
Molecular cloning
Ribosomal protein
Saccharomyces cerevisiae
Thallophyta
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Abstract Article Usage Stats Services MCB Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter current issue Spotlights in the Current Issue MCB About MCB Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy MCB RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 0270-7306 Online ISSN: 1098-5549 Copyright © 2014 by the American Society for Microbiology.   For an alternate route to MCB .asm.org, visit: MCB       
AbstractList In the yeast Saccharomyces cerevisiae, L30 is one of many ribosomal proteins that is encoded by two functional genes. We have cloned and sequenced RPL30B, which shows strong homology to RPL30A. Use of mRNA as a template for a polymerase chain reaction demonstrated that RPL30B contains an intron in its 5' untranslated region. This intron has an unusual 5' splice site, C/GUAUGU. The genomic copies of RPL30A and RPL30B were disrupted by homologous recombination. Growth rates, primer extension, and two-dimensional ribosomal protein analyses of these disruption mutants suggested that RPL30A is responsible for the majority of L30 production. Surprisingly, meiosis of a diploid strain carrying one disrupted RPL30A and one disrupted RPL30B yielded four viable spores. Ribosomes from haploid cells carrying both disrupted genes had no detectable L30, yet such cells grew with a doubling time only 30% longer than that of wild-type cells. Furthermore, depletion of L30 did not alter the ratio of 60S to 40S ribosomal subunits, suggesting that there is no serious effect on the assembly of 60S subunits. Polysome profiles, however, suggest that the absence of L30 leads to the formation of stalled translation initiation complexes.
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In the yeast Saccharomyces cerevisiae, L30 is one of many ribosomal proteins that is encoded by two functional genes. We have cloned and sequenced RPL30B, which shows strong homology to RPL30A. Use of mRNA as a template for a polymerase chain reaction demonstrated that RPL30B contains an intron in its 5' untranslated region. This intron has an unusual 5' splice site, C/GUAUGU. The genomic copies of RPL30A and RPL30B were disrupted by homologous recombination. Growth rates, primer extension, and two-dimensional ribosomal protein analyses of these disruption mutants suggested that RPL30A is responsible for the majority of L30 production. Surprisingly, meiosis of a diploid strain carrying one disrupted RPL30A and one disrupted RPL30B yielded four viable spores. Ribosomes from haploid cells carrying both disrupted genes had no detectable L30, yet such cells grew with a doubling time only 30% longer than that of wild-type cells.
Author D M Baronas-Lowell
J R Warner
AuthorAffiliation Department of Cell Biology, Albert Einstein College of Medicine, Bronx, New York 10461
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Issue 10
Keywords Fungi
Gene
Nucleotide sequence
Ascomycetes
Molecular cloning
Ribosomal protein
Saccharomyces cerevisiae
Thallophyta
Language English
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In the yeast Saccharomyces cerevisiae, L30 is one of many ribosomal proteins that is encoded by two functional genes. We have cloned and sequenced RPL30B,...
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StartPage 5235
SubjectTerms Amino Acid Sequence
Base Sequence
Biological and medical sciences
Blotting, Northern
Cloning, Molecular
DNA Mutational Analysis
Electrophoresis, Gel, Two-Dimensional
Fundamental and applied biological sciences. Psychology
Genes, Fungal
Microbiology
Molecular Sequence Data
Mycology
Polymerase Chain Reaction
Polyribosomes - metabolism
Restriction Mapping
Ribosomal Proteins - physiology
Ribosomes - physiology
Ribosomes - ultrastructure
RNA Splicing
RNA, Viral - genetics
Saccharomyces cerevisiae
Saccharomyces cerevisiae - physiology
Structure-Activity Relationship
Title Ribosomal protein L30 is dispensable in the yeast Saccharomyces cerevisiae
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Volume 10
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