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The synthesis and degradation of ras-related gene products during growth and differentiation in Dictyostelium discoideum
Authors:Gerald Weeks   Tony Pawson
Affiliation:Department of Microbiology. University of British Columbia, Vancouver, B.C. V6T 1W5, Canada
Abstract:A Dictyostelium discoideum protein with an Mr of 23,000 (p23dd-ras) is structurally related to the mammalian proto-oncogene ras-gene product, p21ras, and is specifically precipitated from cell-free extracts of D. discoideum by the Y13-259 monoclonal antibody against p21ras. p23dd-ras was degraded at rates that were very similar to those observed for total protein during both growth and differentiation, suggesting that the previously reported decline in p23dd-ras levels during differentiation is due to a change in the rate of synthesis rather than a change in the rate of degradation. p23dd-ras synthesis did not decrease immediately after the initiation of differentiation, but rather its rate of synthesis increased for the first 1-2 h, suggesting that p23dd-ras is not rapidly down-regulated in response to nutrient deprivation. There were differences in the extent of p23dd-ras turnover during the differentiation of the three tested strains, A-3, NC4, and V12-M2. The relative level of p23dd-ras dropped most rapidly in V12-M2, which may reflect the slightly faster differentiation process exhibited by this strain. In all three strains, very little p23dd-ras was present by the end of the differentiation process. A second protein with an Mr of 24,000 (p24dd-ras) was also immunoprecipitated using the Y13-259 antibody. The amount of p24dd-ras was small or undetectable in vegetative cells, but relatively larger amounts of p24dd-ras were synthesized in pseudoplasmodial cells. We found no evidence to suggest that p24dd-ras is a precursor of p23dd-ras.(ABSTRACT TRUNCATED AT 250 WORDS)
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