|David I. Ratner
Professor of Biology, Amherst College
Ph.D.: Harvard University
Regulation of Gene Expression in Dictyostelium
Embryonic tissues of a multicellular eukaryote diverge through the selective expression of their common genetic constitution. The long-term goal of our research is to understand the basis of cell-specific gene expression during differentiation. Our experimental system is the cellular slime mold, Dictyostelium discoideum, in which spore and stalk cells develop from common vegetative precursors.
One aspect of our research involves the analysis of the function of a developmentally regulated Dictyostelium gene, DG17. DG17 encodes a protein with homology to a small class of zinc finger proteins known as "RING fingers" and to the TRAF family of signal transduction proteins. The function of DG17 is presently unknown but may involve gene regulation. Our approach has been to create DG17 null mutants through gene targeting. We have replaced the DG17 gene of amoebae with an interrupted, inactive copy, but before a phenotype can be determined we must also inactivate a newly discovered DG17 homologue which we have now cloned. As part of these gene targeting experiments, we have investigated the mechanism of chromosomal integration of added DNA and especially the fate of the ends of targeting DNA molecules.
A more open-ended search for components of the transcriptional machinery involves the use of green fluorescent protein (GFP) reporter constructs and fluorescence activated cell sorting, or FACS. We have shown that FACS can purify cells in which GFP has been activated by a Dictyostelium promoter and hope to select developmental mutants altered in signal transduction or transcriptional activation.
Finally, our lab collaborates with several other groups at Amherst College in an analysis of the structure and function of mouse catalytic antibodies.
Pauyo, T. '05, G. J. Hilinski '04, P. T. Chiu '02, D. E. Hansen, Y. J. Choi, D. I. Ratner, N. Shah-Mahoney, C. A. Southern, and P. B. O'Hara (2006). Genetic and fluorescence studies of affinity maturation in related antibodies. Molecular Immunology 43: 812-821.
Tekinay, T., Ennis, H.L., Wu, M.Y., Nelson, M., Kessin, R.H. and Ratner, D.I. 2003. Genetic Interactions of the E3 Ubiquitin Ligase Component FbxA with Cyclic AMP Metabolism and a Histidine Kinase Signaling Pathway During Dictyostelium discoideum Development. Eukaryotic Cell 2 : 618-626.
John D. Bishop, Byoung C. Moon, Faith Harrow, David Ratner, Richard H. Gomer, Robert P. Dottin and Derrick T. Brazill (2002). A second UDP-glucose pyrophosphorylase is required for differentiation and development in Dictyostelium discoideum . Journal of Biological Chemistry 277: 32430-32437.
Nadin, B.M., Mah, C.S., Scharff, J. R. and Ratner, D.I. 2000. The regulative capacity of prespore amoebae as demonstrated by fluorescence-activated cell sorting and green fluorescent protein. Developmental Biology 217 : 173-178.
Wang, Z., Raifu, M.., Howard,M., Smith, L., Hansen, D., Goldsby, R., and Ratner, D. (2000). Universal PCR amplification of mouse immunoglobulin gene variable regions: the design of degenerate primers and an assessment of the effect of DNA polymerase 3' to 5' exonuclease activity, J. Immunological Methods 233: 167-177 .
Nadin, B.M. Mah, C.S., Scharff, J. R. and Ratner, D.I. (2000). The Regulative Capacity of Prespore Amoebae as Demonstrated by Fluorescence-Activated Cell Sorting and Green Fluorescent Protein, Developmental Biology 217: 173-178.
Shah-Mahoney, N., Hampton, T., Vidaver, R. and Ratner, D. (1997) Blocking the ends of transforming DNA enhances gene targeting in Dictyostelium. Gene 203, 33-41.
DeSilver, D.A., Benedict, M.A. and Ratner, D.I. (1991) Effects of protein synthesis inhibition on the transcription and transcript stability of Dictyostelium prespore genes. Biochim. Biophys. Acta. 1089, 309-319.
Smith, S.S. and Ratner, D.I. (1991) Lack of 5-methylcytosine in Dictyostelium discoideum DNA. Biochem. J. 277, 273-275.
Benedict, M.A., DeSilver, D.A., Pelletier, D.E., Pentz, W.H. and Ratner, D.I. (1991) Developmental protein synthesis is required for the transcription of Dictyostelium prespore genes. Dev. Genet. 12, 113-122.
Ratner, D.I., Pentz, W.H. and Pelletier, D.A. (1989) Prespore gene expression in Dictyostelium requires concomitant protein synthesis. Biochim. Biophys. Acta. 1008, 71-78.
Ratner, D. I. (1986). Equivalence of intracellular pH of differentiating Dictyostelium cell types. Nature 321:180-182.