Open AccessA system for the analysis of yeast ribosomal DNA mutations.
Author(s) -
K. Musters,
Jaap Venema,
Gerard van der Linden,
Harm van Heerikhuizen,
J. Klootwijk,
Rudi J. Planta
Publication year - 1989
Publication title -
molecular and cellular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.14
H-Index - 327
eISSN - 1067-8824
pISSN - 0270-7306
DOI - 10.1128/mcb.9.2.551
Subject(s) - biology , ribosomal rna , genetics , ribosomal dna , gene , saccharomyces cerevisiae , operon , eukaryotic large ribosomal subunit , 23s ribosomal rna , plasmid , microbiology and biotechnology , 18s ribosomal rna , ribosome , rna , escherichia coli , phylogenetics
To develop a system for the analysis of eucaryotic ribosomal DNA (rDNA) mutations, we cloned a complete, transcriptionally active rDNA unit from the yeast Saccharomyces cerevisiae on a centromere-containing yeast plasmid. To distinguish the plasmid-derived ribosomal transcripts from those encoded by the rDNA locus, we inserted a tag of 18 base pairs within the first expansion segment of domain I of the 26S rRNA gene. We demonstrate that this insertion behaves as a neutral mutation since tagged 26S rRNA is normally processed and assembled into functional ribosomal subunits. This system allows us to study the effect of subsequent mutations within the tagged rDNA unit on the biosynthesis and function of the rRNA. As a first application, we wanted to ascertain whether the assembly of a 60S subunit is dependent on the presence in cis of an intact 17S rRNA gene. We found that a deletion of two-thirds of the 17S rRNA gene has no effect on the accumulation of active 60S subunits derived from the same operon. On the other hand, deletions within the second domain of the 26S rRNA gene completely abolished the accumulation of mature 26S rRNA.