Hotspots for unselected Ty1 transposition events on yeast chromosome III are near tRNA genes and LTR sequences

A collection of yeast strains bearing single marked Ty1 insertions on chromosome III was generated. Over 100 such insertions were physically mapped by pulsed-field gel electrophoresis. These insertions are very nonrandomly distributed. Thirty-two such insertions were cloned by the inverted PCR technique, and the flanking DNA sequences were determined. The sequenced insertions all fell within a few very limited regions of chromosome III. Most of these regions contained tRNA coding regions and/or LTRs of preexisting transposable elements. Open reading frames were disrupted at a far lower frequency than expected for random transposition. The results suggest that the Ty1 integration machinery can detect regions of the genome that may represent "safe havens" for insertion. These regions of the genome do not contain any special DNA sequences, nor do they behave as particularly good targets for Ty1 integration in vitro, suggesting that the targeted regions have special properties allowing specific recognition in vivo.

[1]  R. Craigie Hotspots and warm spots: integration specificity of retroelements. , 1992, Trends in genetics : TIG.

[2]  D. Hartl,et al.  Genetic applications of an inverse polymerase chain reaction. , 1988, Genetics.

[3]  O H Jarrett Retroviruses. , 1987, Archives of disease in childhood.

[4]  P. Philippsen,et al.  Preferential integration of yeast transposable element Ty into a promoter region , 1984, Nature.

[5]  G. Natsoulis,et al.  Doubling Ty1 element copy number in Saccharomyces cerevisiae: host genome stability and phenotypic effects. , 1991, Genetics.

[6]  R. W. Davis,et al.  Studies on the transposable element Ty1 of yeast. I. RNA homologous to Ty1. II. Recombination and expression of Ty1 and adjacent sequences. , 1981, Cold Spring Harbor symposia on quantitative biology.

[7]  P. Brown,et al.  Correct integration of retroviral DNA in vitro , 1987, Cell.

[8]  J. Coffin,et al.  Highly preferred targets for retrovirus integration , 1988, Cell.

[9]  H. Varmus,et al.  Retroviral integration into minichromosomes in vitro. , 1992, The EMBO journal.

[10]  R. Anwar,et al.  A 'hot-spot' for Ty transposition on the left arm of yeast chromosome III. , 1986, Nucleic acids research.

[11]  J. Boeke,et al.  A specific terminal structure is required for Ty1 transposition. , 1990, Genes & Development.

[12]  G. Fink,et al.  A general method for the chromosomal amplification of genes in yeast. , 1988, Science.

[13]  H. Feldmann,et al.  Different patterns of transposable elements in the vicinity of tRNA genes in yeast: a possible clue to transcriptional modulation. , 1985, Biological chemistry Hoppe-Seyler.

[14]  K. Isono,et al.  Chromosome III of Saccharomyces cerevisiae: An ordered clone bank, a detailed restriction map and analysis of transcripts suggest the presence of 160 genes , 1990, Yeast.

[15]  G. Natsoulis,et al.  Ty1 transposition in Saccharomyces cerevisiae is nonrandom. , 1989, Genetics.

[16]  G. Fink,et al.  Ty-mediated gene expression of the LYS2 and HIS4 genes of Saccharomyces cerevisiae is controlled by the same SPT genes. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[17]  J. Boeke,et al.  Inhibition of Ty1 transposition by mating pheromones in Saccharomyces cerevisiae , 1991, Molecular and cellular biology.

[18]  C. Newlon,et al.  Polymorphisms on the right arm of yeast chromosome III associated with Ty transposition and recombination events. , 1987, Nucleic acids research.

[19]  N. Kleckner,et al.  A symmetrical six-base-pair target site sequence determines Tn10 insertion specificity , 1982, Cell.

[20]  B. Dujon,et al.  The complete DNA sequence of yeast chromosome III , 1992, Nature.

[21]  J. Boeke,et al.  High-frequency deletion between homologous sequences during retrotransposition of Ty elements in Saccharomyces cerevisiae. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[22]  G. Fink,et al.  Ty elements transpose through an RNA intermediate , 1985, Cell.

[23]  C. Newlon,et al.  Analysis of a circular derivative of Saccharomyces cerevisiae chromosome III: a physical map and identification and location of ARS elements. , 1991, Genetics.

[24]  J. Boeke,et al.  4 Yeast Transposable Elements , 1991 .

[25]  H. Feldmann,et al.  Ty4, a novel low-copy number element in Saccharomyces cerevisiae: one copy is located in a cluster of Ty elements and tRNA genes. , 1989, Nucleic acids research.

[26]  Ronald W. Davis,et al.  Reversion of a promoter deletion in yeast , 1982, Nature.

[27]  T. Cooper,et al.  Tau, sigma, and delta. A family of repeated elements in yeast. , 1984, The Journal of biological chemistry.

[28]  H. Feldmann,et al.  Ty1 and delta elements occur adjacent to several tRNA genes in yeast. , 1982, The EMBO journal.

[29]  D. Chalker,et al.  Ty3 integrates within the region of RNA polymerase III transcription initiation. , 1992, Genes & development.

[30]  R. D. Gietz,et al.  Elimination of the yeast RAD6 ubiquitin conjugase enhances base-pair transitions and G.C----T.A transversions as well as transposition of the Ty element: implications for the control of spontaneous mutation. , 1992, Genetics.

[31]  Daniel F. Voytas,et al.  Yeast retrotransposon revealed , 1992, Nature.

[32]  M. Olson,et al.  Insertion of a repetitive element at the same position in the 5'-flanking regions of two dissimilar yeast tRNA genes. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[33]  Robert Craigie,et al.  The IN protein of Moloney murine leukemia virus processes the viral DNA ends and accomplishes their integration in vitro , 1990, Cell.

[34]  R. W. Davis,et al.  Separation of large DNA molecules by contour-clamped homogeneous electric fields. , 1986, Science.

[35]  E. Dubois,et al.  Mating signals control expression of mutations resulting from insertion of a transposable repetitive element adjacent to diverse yeast genes , 1980, Cell.

[36]  C. Newlon,et al.  Nucleotide sequence characterization of Ty 1-17, a class II transposon from yeast. , 1985, Nucleic acids research.

[37]  M V Olson,et al.  Physical map of the Saccharomyces cerevisiae genome at 110-kilobase resolution. , 1991, Genetics.

[38]  D. Garfinkel,et al.  Transposon tagging using Ty elements in yeast. , 1988, Genetics.

[39]  F. Bushman,et al.  Retroviral DNA integration directed by HIV integration protein in vitro. , 1990, Science.

[40]  D. Chalker,et al.  Transfer RNA genes are genomic targets for de Novo transposition of the yeast retrotransposon Ty3. , 1990, Genetics.

[41]  M. Culbertson,et al.  The yeast frameshift suppressor gene SUF16-1 encodes an altered glycine tRNA containing the four-base anticodon 3'-CCCG-5'. , 1982, Gene.

[42]  S. Liebman,et al.  Analysis of yeast retrotransposon Ty insertions at the CAN1 locus. , 1989, Genetics.

[43]  J. Coffin,et al.  Nonrandom integration of retroviral DNA in vitro: effect of CpG methylation. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[44]  Harold E. Varmus,et al.  Nucleosomes, DNA-binding proteins, and DNA sequence modulate retroviral integration target site selection , 1992, Cell.

[45]  P. Morcos,et al.  Sigma elements are position-specific for many different yeast tRNA genes. , 1988, Nucleic acids research.

[46]  Fred Winston,et al.  Methods in Yeast Genetics: A Laboratory Course Manual , 1990 .

[47]  R. Lathe,et al.  Linker tailing: unphosphorylated linker oligonucleotides for joining DNA termini. , 1984, DNA.

[48]  M. Olson,et al.  Physical maps of the six smallest chromosomes of Saccharomyces cerevisiae at a resolution of 2.6 kilobase pairs. , 1993, Genetics.

[49]  S. Antonarakis,et al.  Methods for cloning large DNA segments as artificial chromosomes in S. cerevisiae. , 1990 .

[50]  S. Dutcher,et al.  Internuclear transfer of genetic information in kar1-1/KAR1 heterokaryons in Saccharomyces cerevisiae , 1981, Molecular and cellular biology.