fungicides, protein synthesis, inhibitors, two dimensional gel electrophoresis, cell cycle, prions, alcohol elongation factor 3
My long term interests are on the effects of toxic compounds and on mixtures on macromolecular metabolism. My objective of this study is to define the biochemical effects of toxic compounds and mixtures on macromolecular metabolism at the cellular level. Two dimensional (2D) gel electrophoresis of proteins is used to define the impact of these toxic compounds on metabolism. Computer analysis of 2D gel electrophoresis of protein has advanced to the point where a database can be constructed that will contain information on how a specific toxic compound affects the 3000 to 5000 different proteins synthesized by an individual cell. My goal is to characterize each toxic compound by its effect on the pattern of proteins made by the cell. In turn the pattern of proteins made by the cell under conditions of chemical stress can be used to identify the stress inducing chemical agent. We will use this information to develop a classification of toxic agents based on their overall impact on metabolism expressed in the synthesis of individual proteins. This classification will allow us to examine mixtures of toxic compounds to determine the dominant biochemical effect and to determine whether compounds of this mixture demonstrate antagonistic or synergistic properties.
The other long term research interest in my laboratory concerns the control of cell division studied by 2D gel electrophoresis. Yeast represents an ideal organism to approach the control of cell division at the biochemical level. The basic aspects of yeast cell cycle are well defined in molecular terms due in part to our efforts. A useful collection of temperature sensitive mutants that affect the cell cycle were isolated by Dr. Leland Hartwell and myself some years ago. Selection of different stages in the cell cycle using elutriation is highly precise. Our results suggest that most housekeeping proteins are synthesized continuously while at least some proteins that have critical periodic roles in cell division are synthesized periodically. We are also attempting to identify the proteins encoded by the genes that affect cell division. This identification relies on the translation of isolated mRNA species using our homologous in vitro translation system.
Latter, G.I., Boutell, T., Monardo, P.J., Kobayashi, R., Futcher, B., McLaughlin, C.S. and Garrels, J.I., A Saccharomyces cerevisiae Internet Protein Resource Is Now Available. Electrophoresis, 16: 1003-1007 (1996).
Garrels, J.I., Futcher, B., Kobayashi, R., Latter, G.I., Schwender, B., Volpe, T., Warner, J.R., and McLauglin, C.S. Protein Identifications for a Saccharomyces cerevisiae Protein Database. Electrophoresis, 15: 1466-1486 (1994).
Santangelo, G. M., Tornow, J., McLaughlin, C.S., and Moldave K. Screening A Yeast Promoter Library Leads to the Isolation of the RP29/L32 and SNR17B/RPL37A Divergent Promoters and the discovery of a Gene Encoding Ribosomal Protein L37. Gene 105: 137-138 (1991).
Qin, S., Xie, A., Bonato, C.M., and McLaughlin, C.S. Sequence Analysis of the Translational Elongation Factor 3 from Saccharomyces cerevisae. J. Biol. Chem. 265: 1903-1912 (1990).
Santos, M., Colhurst, D.R., Wills, N., McLaughlin, C.S., and Tuite, M.F. Efficient Translation of the UAG Termination Codon in Candida Species. Current Genetics 17: 487-491 (1990.)