The DNA intermediate in yeast Ty1 element transposition copurifies with virus-like particles: cell-free Ty1 transposition.
Yeast Ty1 elements are retrotransposons that transpose via an RNA intermediate found in a virus-like particle (Ty-VLP). A Ty-encoded reverse transcriptase activity found inside the particles is capable of giving rise to full-length reverse transcripts. The predominant form of these reverse transcripts is a full-length linear duplex DNA. We have developed a cell-free system for transposition of Ty1 DNA molecules into a bacteriophage lambda target. Purified Ty-VLPs and target DNA are the only macromolecular components required for the transposition reaction. A TYB-encoded protein, p90-TYB, contains amino acid sequences that are similar to those of retroviral integrase proteins. Mutations in the integrase coding region abolish transposition both in vivo and in vitro.
Eichinger
D J
DJ
Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
Boeke
J D
JD
D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D010582:Bacteriophage lambda / Q000235:genetics; D001483:Base Sequence; D002458:Cell Fractionation; D002474:Cell-Free System; D002499:Centrifugation, Density Gradient; D004254:DNA Nucleotidyltransferases / Q000235:genetics; D004251:DNA Transposable Elements; D004270:DNA, Circular / Q000302:isolation & purification; D004271:DNA, Fungal / Q000235:genetics* / Q000302:isolation & purification; D004274:DNA, Recombinant / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D019426:Integrases; D009154:Mutation; D010957:Plasmids; D012190:Retroviridae / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid
D000911:Antibodies, Monoclonal; D004251:DNA Transposable Elements; D004270:DNA, Circular; D004271:DNA, Fungal; D004274:DNA, Recombinant; D004254:DNA Nucleotidyltransferases; D019426:Integrases