Olivier_to_post - Olivier et al. A Drosophila SH2-SH3...

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Unformatted text preview: Olivier et al. A Drosophila SH2-SH3 adapter protein implicated in coupling the sevenless tyrosine kinase to an activator of Ras guanine nucleotide exchange, Sos A. Major Questions The "big picture" in this paper is very similar to last paper. The authors want to discover components that act downstream of a receptor tyrosine kinase (RTK) during signal transduction. RTKs are important for cell growth (EGF receptor) differentiation decisions during development e.g., Boss in R7 photoreceptor differentiation in y eye Last paper - how is RTK activation linked to activation of PLC ? This paper - how is RTK activation linked to activation of Ras (using Drosophila system to study) Ras Already known Ras (small GTP binding protein (MBOC 15-50) ) becomes activated when RTKs are activated Blocking Ras activity blocks proliferative effects of RTK signaling blocks MAP kinase activity Downstream of Ras: Ras activation GEF But HOW does Ras get activated by a RTK? Raf Idea of adapter proteins Already known that SH2 protein domain recognizes phosphotyrosine and is used downstream of RTKs in C. elegans and in mammalian cells (MBOC 15-49) . note: PLC- from last paper was later demonstrated to interact with the autophosphorylated tyrosines in the EGF receptor via SH2 domains in PLC- . Crystal structure: SH2 domain B. Hypothesis: An SH2-domain-containing protein is expected to act downstream of RTKs in Drosophila . What assumption do the authors make to search for this protein? How do they look for it? C. Approach: Molecular Biology, Genetics and Biochemistry Experimental Strategy to identify proteins acting downstream of RTK: D. Tools needed for these experiments Note the use of a Drosophila "cDNA library": When do you use a cDNA library and when do you use a genomic library? mRNA > cDNA > cDNA library: library will be enriched in protein encoding sequences that are expressed in the cells that served as the source of material. Genomic DNA > genomic library: library will contain upstream regulatory regions, introns and other nontranscribed sequence as well as protein-coding sequences. E. Determining the function of "drk - expression in Fies 1. Where is drk expressed?...
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This note was uploaded on 10/19/2009 for the course B M B 460 taught by Professor Hanna-rose,w during the Spring '08 term at Pennsylvania State University, University Park.

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Olivier_to_post - Olivier et al. A Drosophila SH2-SH3...

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