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. Integrati...

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Bibliographic Details
Published in:Science (American Association for the Advancement of Science) 1995-03, Vol.267 (5203), p.1488-1491
Main Authors: Kirchner, Jacqueline, Connolly, Charles M., Sandmeyer, Suzanne B.
Format: Article
Language:English
Subjects:
Biological and medical sciences
DNA
DNA Nucleotidyltransferases - metabolism
Fundamental and applied biological sciences. Psychology
Genes
Genetic mutation
Genetics
Integrases
Literary Devices
Microbiology
Models, Genetic
Mutagenesis, Site-Directed
Plasmids
Polymerase chain reaction
Proteins
Recombination, Genetic
Region of integration
Retroelements
Retroviridae
Ribonucleic acid
RNA
RNA Polymerase III - genetics
RNA, Transfer, Tyr - genetics
Saccharomyces cerevisiae
Saccharomyces cerevisiae - genetics
Site Selection
Transcription Factor TFIIIB
Transcription factors
Transcription Factors - metabolism
Transcription Factors, TFIII
Transcription, Genetic
Transfer RNA
Virology
Virus Integration
Yeast
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Summary: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 extract, and a transcriptionally competent tRNA gene. Use of individual 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.
ISSN:0036-8075
1095-9203
DOI:10.1126/science.7878467