Yeast tRNA ligase mutants are nonviable and accumulate tRNA splicing intermediates.
We show here that yeast tRNA ligase protein is essential in the cell and participates in joining together tRNA half-molecules resulting from excision of the intron by the splicing endonuclease. A haploid yeast strain carrying a chromosomal deletion of the ligase gene is viable only if ligase protein can be supplied from a plasmid copy of the gene. When synthesis of the plasmid-borne ligase gene is repressed, cells eventually die and accumulate endonuclease cut but unligated half-molecules and intervening sequences. Half-molecules that accumulate appear to be fully end-processed. Two temperature-sensitive ligase mutant strains have been isolated; these strains accumulate a similar set of unligated half-molecules at the nonpermissive temperature.
Phizicky
E M
EM
Department of Biochemistry, University of Rochester School of Medicine and Dentistry, New York 14642.
Consaul
S A
SA
Nehrke
K W
KW
Abelson
J
J
D002872:Chromosome Deletion; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D009154:Mutation; D010957:Plasmids; D011130:RNA Ligase (ATP) / Q000235:genetics*; D012326:RNA Splicing; D012343:RNA, Transfer / Q000235:genetics*; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D013696:Temperature
D012343:RNA, Transfer; D011130:RNA Ligase (ATP)