线虫发育及基因功能研究课件

Click to edit Master title style,Click to edit Master text styles,Second level,Third level,Fourth level,Fifth level,#,R7,R8,WT,sevenless,R7 receptor is required for UV response,sev,Boss,R7R8WTsevenlessR7 receptor is,1,Strategy:moving from RTK down:discovering SOS,RAS,GDP,RAS,GTP,Pi,GTP,GDP,GAP,target protein,Active state,Inactive,RTK,Strategy:moving from RTK down,2,Son of Sevenless:hyperactive mutation,Roger and Benejee,Cell 1990,The story about Ron Roger,-,+,Sev/RTK(lf)Sevenless,Sev(lf)+,SOS(gf),R7 fine,Sev,SOS,R7 fate,Leads to Model:,A:yes,B:may not.,Son of Sevenless:hyperactive,3,Drosophila:F1 screen vs F2 screen,X,*,*,X,*,*,F3 homozygotes,mutagen,X,TM,TM,*,*,F1,F2 screen,mutagen,X,*,*,F1,F1 screen,TM,A conversation with Rubin,Drosophila:F1 screen vs F2 s,4,Mike Simon:landmark modify screen,Simon et al.1991 Cell,Sev,RAS,SOS,R7,Rough eye,no R7,Mostly wt,small%defects,Rough eye,Rough eye,High T,Low T,Low T,Low T,Ts allele,X,*,*,F1,F1 screen,TM,+/-,+/-,Mike Simon:landmark modify s,5,SOS encodes an exchange factor for GTPase,Simon et al.1991 Cell,RAS,GDP,RAS,GTP,Pi,GTP,GDP,SOS,GAP,target protein,Active state,Inactive,RTK,SOS encodes an exchange factor,6,线虫发育及基因功能研究课件,7,Moving from receptor down:biochemistry,RAS,GDP,RAS,GTP,Pi,GTP,GDP,GAP,target protein,Active state,Inactive,RTK,SOS,Moving from receptor down:bio,8,Discovery of GRB2,EGFR,Y,EGFR,Ligand activation,Y-,P,Screen,l,GT11 protein expression library,9 GRB proteins,Which one is one mediated Ras activation?,Y-,P,Discovery of GRB2EGFRYEGFRLig,9,Discovery of GRB2:complementary works by biochemists and geneticists,A cDNA clone encoding a novel,widely expressed protein(called growth factor receptor-bound protein 2 or GRB2)containing one src homology 2(SH2)domain and two SH3 domains was isolated.,Immunoblotting experiments indicate that GRB2 associates with tyrosine-phosphorylated epidermal growth factor receptors(EGFRs)and platelet-derived growth factor receptors(PDGFRs),via its SH2 domain.Interestingly,GRB2 exhibits striking structural and functional homology to the C.elegans protein sem-5.It has been shown that sem-5 and two other genes called let-23(EGFR like)and let-60(ras like)lie along the same signal transduction pathway controlling C.elegans vulval induction,.To examine whether GRB2 is also a component of ras signaling in mammalian cells,microinjection studies were performed.While injection of GRB2 or H-ras proteins alone into quiescent rat fibroblasts did not have mitogenic effect,microinjection of GRB2 together with H-ras protein stimulated DNA synthesis.These results suggest that GRB2/sem-5 plays a crucial role in a highly conserved mechanism for growth factor control of ras signaling.,Lowenstein et al.Cell.,Aug,.1992,Discovery of GRB2:complement,10,The sem-5 story-early 1992,SEM-5 was identified as by a suppressor mutant screen,Clark et al.Nature,Mar,1992,lin-15(-),Multivulva,sem-5(-)Vulvaless,lin-15(-);,sem-5(-)Vulvaless,Ras,SEM-5,LIN-15,EGFR,LIN-15,Ras,Vulval induction,Known,Since 1990,SEM-5 is homologous to GRB-2.Combine the data from the two field:,Ras,GRB2/SEM-5,EGFR,ras(gf),Mutivulva,sem-5(-)Vulvaless,ras(gf);,sem-5(-),Multivulva,The sem-5 story-early 1992SE,11,The rest of the story is there in a rush and clash-mid 1993,Olivier JP,Raabe T,Henkemeyer M,Dickson B,Mbamalu G,Margolis B,Schlessinger J,Hafen E,Pawson.A Drosophila SH2-SH3 adaptor protein implicated in coupling the sevenless tyrosine kinase to an activator of Ras guanine nucleotide exchange,Sos.,Cel,l.1993 Apr 9;73:179-91.,Egan SE,Giddings BW,Brooks MW,Buday L,Sizeland AM,Weinberg RA.,Association of Sos Ras exchange protein with Grb2 is implicated in tyrosine kinase signal transduction and transformation.,Nature,.1993 May 6;363:45-51.,Rozakis-Adcock M,Fernley R,Wade J,Pawson T,Bowtell D.,The SH2 and SH3 domains of mammalian Grb2 couple the EGF receptor to the Ras activator mSos1.,Nature,.1993 May 6;363:83-5.,Buday L,Downward J.,Epidermal growth factor regulates p21ras through the formation of a complex of receptor,Grb2 adapter protein,and Sos nucleotide exchange factor.,Cell,.1993 May 7;73:611-20.,Chardin P,Camonis JH,Gale NW,van Aelst L,Schlessinger J,Wigler MH,Bar-Sagi D.,Human Sos1:a guanine nucleotide exchange factor for Ras that binds to GRB2.,Scienc,e.1993 May 28;260:1338-43.,Skolnik EY,Batzer A,Li N,Lee CH,Lowenstein E,Mohammadi M,Margolis B,Schlessinger J.,The function of GRB2 in linking the insulin receptor to Ras signaling pathways.,Science,.1993 Jun 25;260:1953-5.,Baltensperger K,Kozma LM,Cherniack AD,Klarlund JK,Chawla A,Banerjee U,Czech MP.,Binding of the Ras activator son of sevenless to insulin receptor substrate-1 signaling complexes.,Science,.1993 Jun 25;260:1950-2.,Simon MA,Dodson GS,Rubin GM.,SH3-SH2-SH3 protein is required for p21Ras1 activation and binds to sevenless and Sos proteins in vitro.,Cel,l.1993 Apr 9;73:169-77.,The rest of the story is there,12,Many Small GTPases have a conserved C-terminus,Many Small GTPases have a cons,13,Ras is lipid modified for i