The Rad27 (Fen-1) nuclease inhibits Ty1 mobility in Saccharomyces cerevisiae.
Although most Ty1 elements in Saccharomyces cerevisiae are competent for retrotransposition, host defense genes can inhibit different steps of the Ty1 life cycle. Here, we demonstrate that Rad27, a structure-specific nuclease that plays an important role in DNA replication and genome stability, inhibits Ty1 at a post-translational level. We have examined the effects of various rad27 mutations on Ty1 element retrotransposition and cDNA recombination, termed Ty1 mobility. The point mutations rad27-G67S, rad27-G240D, and rad27-E158D that cause defects in certain enzymatic activities in vitro result in variable increases in Ty1 mobility, ranging from 4- to 22-fold. The C-terminal frameshift mutation rad27-324 confers the maximum increase in Ty1 mobility (198-fold), unincorporated cDNA, and insertion at preferred target sites. The null mutation differs from the other rad27 alleles by increasing the frequency of multimeric Ty1 insertions and cDNA recombination with a genomic element. The rad27 mutants do not markedly alter the levels of Ty1 RNA or the TyA1-gag protein. However, there is an increase in the stability of unincorporated Ty1 cDNA in rad27-324 and the null mutant. Our results suggest that Rad27 inhibits Ty1 mobility by destabilizing unincorporated Ty1 cDNA and preventing the formation of Ty1 multimers.
Sundararajan
Anuradha
A
Gene Regulation and Chromosome Biology Laboratory, National Cancer Institute, Frederick, Maryland 21702, USA.
Lee
Bum-Soo
BS
Garfinkel
David J
DJ
D004706:Endodeoxyribonucleases / Q000235:genetics / Q000378:metabolism*; D045585:Flap Endonucleases; D009154:Mutation; D011995:Recombination, Genetic; D018626:Retroelements; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins
D018626:Retroelements; D029701:Saccharomyces cerevisiae Proteins; D004706:Endodeoxyribonucleases; D045585:Flap Endonucleases; C468323:RAD27 protein, S cerevisiae