Sulfite mutants representing five complementation groups, previously derived from
an ethyl methanesulfonate-treated haploid strain of Saccharomyces cerevisiae were studied.
Although the wildtype S. cerevisiae strain used (isogenic to X2180-1 A) had a basal
tolerance for sulfite (7 μM free H₂SO₃), the sensitive and resistant mutants were found to
tolerate less than 3 to 5.5, or greater than 19 μM free H₂SO₃, respectively. No apparent
correlation was found between the response to sulfite and generation time in rich (YEPD)
or minimal media. Resistant mutant 11-1 had an extended lag phase relative to wildtype.
Mutant and wildtype proteins were labeled with ³⁵S-methionine to determine differences in
banding patterns due to sulfite-specific induction or disappearance of polypeptides. No
obvious differences following SDS-PAGE and autoradiography were observed upon
induction with 0.213 μM free H₂SO₃. No consistent correlations were found between the
sulfite phenotypes and responses to other reducing agents. Sensitive mutant 35-2 appeared
to be three to ten times more sensitive to dithiothreitol than wildtype and sensitive mutant
47-9 was three to four times more sensitive to sodium nitrite and three to seven times more
sensitive to sodium thiosulfate than wildtype. Log phase cells of sensitive mutant 33-2
were found to have significantly less glutathione than wildtype. Wildtype contained 62.6 nmol min⁻¹ mg protein⁻¹ (62.6 mU mg protein⁻¹) glutathione reductase (GR) and 2.78
nmol min⁻¹ mg protein⁻¹ (2.78 mU mg protein⁻¹) glutathione S-transferase (GST). Log
phase cells of one resistant mutant showed a significantly higher level of GR than wildtype,
135%. The resistant mutants as well as some of the sensitive mutants had reduced GST
levels. Survival rates of the mutants in buffer in the presence of sulfite did not correlate
with their sensitive or resistant phenotypes, suggesting that survival and growth in the
presence of sulfite are not necessarily related functions. Relative to wildtype, survival
upon prolonged storage at 4°C was markedly reduced for two of the four sensitive
mutants, one of which was 33-2, and was enhanced for one resistant and another sensitive
mutant. / Graduation date: 1992
| Identifer | oai:union.ndltd.org:ORGSU/oai:ir.library.oregonstate.edu:1957/27172 |
| Date | 18 March 1992 |
| Creators | Wightman, JoLynne Dee |
| Contributors | Bakalinsky, Alan Tagore |
| Source Sets | Oregon State University |
| Language | en_US |
| Detected Language | English |
| Type | Thesis/Dissertation |
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