Replication infidelity during a single cycle of Ty1 retrotransposition.
Retroviruses undergo a high frequency of genetic alterations during the process of copying their RNA genomes. However, little is known about the replication fidelity of other elements that transpose via reverse transcription of an RNA intermediate. The complete sequence of 29 independently integrated copies of the yeast retrotransposon Ty1 (173,043 nt) was determined, and the mutation rate during a single cycle of replication was calculated. The observed base substitution rate of 2.5 x 10(-5) bp per replication cycle suggests that this intracellular element can mutate as rapidly as retroviruses. The pattern and distribution of errors in the Ty1 genome is nonrandom and provides clues to potential in vivo molecular mechanisms of reverse transcriptase-mediated error generation, including heterogeneous RNase H cleavage of Ty1 RNA, addition of terminal nontemplated bases, and transient dislocation and realignment of primer-templates. Overall, analysis of errors generated during Ty1 replication underscores the utility of a genetically tractable model system for the study of reverse transcriptase fidelity.
Gabriel
A
A
Department of Molecular Biology and Biochemistry, Rutgers University, Piscataway, NJ 08855, USA.
Willems
M
M
Mules
E H
EH
Boeke
J D
JD
D001483:Base Sequence; D004261:DNA Replication; D004271:DNA, Fungal / Q000235:genetics; D004926:Escherichia coli; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009154:Mutation; D009690:Nucleic Acid Conformation; D010957:Plasmids; D012331:RNA, Fungal / Q000096:biosynthesis / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D012091:Repetitive Sequences, Nucleic Acid; D018626:Retroelements; D016914:Ribonuclease H / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013698:Templates, Genetic
D004271:DNA, Fungal; D012331:RNA, Fungal; D018626:Retroelements; D012194:RNA-Directed DNA Polymerase; D016914:Ribonuclease H