Self-Perpetuating Structural States in Biology, Disease, and Genetics (2002) / Chapter Skim
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Changes in the middle region of Sup35 profoundly alter the nature of epigenetic inheritance for the yeast prion [PSI+]
Changes in the middle region of Sup35 profoundly alter the nature of epigenetic inheritance for the yeast prion [PSI+]
Pages 70-77
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From page 70... ...
N regions from even distantly related Hemiascomycetes are rich in glut amine and asparagine residues (16, 19~. M regions are highly charged, and their sequences are heavily biased toward a subset of charged amino acids (9, 16, 18, 19~.
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From page 71... ...
Primers were as follows: 5'-CCA CAG TCT CAA GGT AAG GAT GGT AAG GAT GGT AAG GAT GGT AAG GAT GGT AAG GAT GGT AAG GAT GGT TTT GGT GGT AAA OAT-3' and 5'-ATC TTT ACC ACC AAA ACC ATC CTT ACC ATC CTT ACC ATC CTT ACC ATC CTT ACC ATC CTT ACC ATC CTT ACC TTG AGA CTG TGG-3'. The SUP35 N-KDG6-C fragment was then PCR amplified and inserted into the SUP35 integrative construct pJLI-SUP35 at the EcoRI/BamHI sites.
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This was confirmed by 4:0 segregation of the suppressor phenotype in crosses to WT tpsi-] cells and curing by growth on medium containing Gdn HCl (data not shown)
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The Human Topoisomerase Linker Restores Mitotic Stability but Causes Meiotic Instability. The human topoisomerase linker (T)
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strains, including a suppressor phenotype that was eliminated by plating to media containing Gdn HCl (see Table 1~. It also showed strong mitotic stability.
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and [PSI+] strains containing the linker from human topoisomerase I in place of the M linker of Sup35p were plated onto YPD; [PSI+]
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phenotype and the Sup35N9C protein returned to the soluble state. Thus, Sup35N9Cp can participate in a heritable phenotypic change caused by a protein-only mechanism that exhibits Mendelian segregation, a striking departure from ordinary prion behavior.
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From page 77... ...
(1994) Methods in Yeast Genetics: A Laboratory Manual (Cold Spring Harbor Lab.
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