TY - JOUR
T1 - In-frame linker insertion mutagenesis of yeast transposon Ty1
T2 - phenotypic analysis
AU - Braiterman, L. T.
AU - Monokian, G. M.
AU - Eichinger, D. J.
AU - Merbs, S. L.
AU - Gabriel, A.
AU - Boeke, J. D.
N1 - Funding Information:
J. Curcio, D. Garfinkel, M. Kupiec and G. Sharon kindly provided unpublished data on the behavior of IN mutants prior to publication. We thank D. Moore and E. Caputo for technical assistance and S. Devine for helpful comments on the manuscript. This work was supported in part by NSF predoctoral training grant RCD-9154644 (L.T.B.), Public Health Service postdoctoral training grant CA-09139 from the National Institutes of Health, Public Health Service grant GM-07445 from the National Institutes of Health (S.L.M.), and a grant from the Lucille P. Markey Charitable Trust (A.G.). Supported in part by Public Health Service grant GM-36481 from the National Institutes of Health and American Cancer Society Faculty Research Award FRA-366 (J.D.B.).
PY - 1994/2/11
Y1 - 1994/2/11
N2 - A plasmid bearing a transpositionally functional GAL1::Tyl fusion was mutagenized by insertion of four or five codons semirandomly throughout the plasmid. This collection of mutant plasmids was introduced into yeast cells and studied with regard to the properties of the mutant Ty1-encoded proteins and the transposition phenotypes observed. All of the transposition-inactivating mutations were previously found to be recessive with the exception of a single mutation in TYA. In this mutant, TYA protein of normal abundance is produced, but the virus-like particles containing this protein are unstable and have aberrant behavior. The effects of mutations in noncoding regions, as well as the capsid protein coding region TYA, and the regions encoding the protease, integrase and reverse transcriptase proteins are described. Effects on gene expression, types of proteins produced, proteolysis of precursor proteins, virus-like particle structure, and biochemical activities of the encoded proteins are summarized. In addition, we show that one of the mutations in the 3' LTR represents a new nonessential site into which foreign marker DNA can be inserted without compromising transposition.
AB - A plasmid bearing a transpositionally functional GAL1::Tyl fusion was mutagenized by insertion of four or five codons semirandomly throughout the plasmid. This collection of mutant plasmids was introduced into yeast cells and studied with regard to the properties of the mutant Ty1-encoded proteins and the transposition phenotypes observed. All of the transposition-inactivating mutations were previously found to be recessive with the exception of a single mutation in TYA. In this mutant, TYA protein of normal abundance is produced, but the virus-like particles containing this protein are unstable and have aberrant behavior. The effects of mutations in noncoding regions, as well as the capsid protein coding region TYA, and the regions encoding the protease, integrase and reverse transcriptase proteins are described. Effects on gene expression, types of proteins produced, proteolysis of precursor proteins, virus-like particle structure, and biochemical activities of the encoded proteins are summarized. In addition, we show that one of the mutations in the 3' LTR represents a new nonessential site into which foreign marker DNA can be inserted without compromising transposition.
KW - Retrotransposons
KW - Saccharomyces cerevisiae
KW - codon insertion mutations
KW - integrase
KW - protease
KW - reverse transcriptase
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U2 - 10.1016/0378-1119(94)90518-5
DO - 10.1016/0378-1119(94)90518-5
M3 - Article
C2 - 8112584
AN - SCOPUS:0028211989
SN - 0378-1119
VL - 139
SP - 19
EP - 26
JO - Gene
JF - Gene
IS - 1
ER -