Requirement of RNA polymerase III transcription factors for in vitro position-specific integration of a retroviruslike element.
The yeast retroviruslike element Ty3 inserts at the transcription initiation sites of genes transcribed by RNA polymerase III (Pol III). An in vitro integration assay was developed with the use of Ty3 viruslike particles and a modified SUP2 tyrosine transfer RNA (tRNA(Tyr)) gene target. Integration was position-specific and required Ty3 integrase, Pol III transcription factor (TF) IIIB-, TFIIIC-, and Pol III-containing fractions showed that TFIIIB and TFIIIC, together, were sufficient for position-specific Ty3 integration, but not for transcription. This report demonstrates that in vitro integration of a retroelement can be targeted by cellular proteins.
Kirchner
J
J
Department of Microbiology and Molecular Genetics, University of California, Irvine 92717.
Connolly
C M
CM
Sandmeyer
S B
SB
D004254:DNA Nucleotidyltransferases / Q000378:metabolism; D019426:Integrases; D008957:Models, Genetic; D016297:Mutagenesis, Site-Directed; D012320:RNA Polymerase III / Q000235:genetics*; D012365:RNA, Transfer, Tyr / Q000235:genetics; D011995:Recombination, Genetic; D018626:Retroelements; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D035361:Transcription Factor TFIIIB; D014157:Transcription Factors / Q000378:metabolism*; D019474:Transcription Factors, TFIII; D014158:Transcription, Genetic; D016662:Virus Integration; D000098832:Transcription Factor TFIIIC
D012365:RNA, Transfer, Tyr; D018626:Retroelements; D035361:Transcription Factor TFIIIB; D014157:Transcription Factors; D019474:Transcription Factors, TFIII; D000098832:Transcription Factor TFIIIC; D004254:DNA Nucleotidyltransferases; D019426:Integrases; D012320:RNA Polymerase III