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S Li P Janosch M Tanji G C Rosenfeld J C Waymire H Mischak W Kolch J M Sedivy 《The EMBO journal》1995,14(4):685-696
We have identified the beta (beta) isoform of the 14-3-3 family of proteins as an activator of the Raf-1 protein kinase. 14-3-3 was isolated in a yeast two-hybrid screen for Raf-1 kinase domain binding proteins. Purified bovine brain 14-3-3 interacted specifically with both c-Raf-1 and the isolated Raf-1 kinase domain. Association was sensitive to the activation status of Raf-1; 14-3-3 bound to unactivated Raf-1, but not Raf-1 activated by protein kinase C alpha or Ras and Lck. The significance of these interactions under physiological conditions was demonstrated by co-immunoprecipitation of Raf-1 and 14-3-3 from extracts of quiescent, but not mitogen-stimulated, NIH 3T3 cells. 14-3-3 was not a preferred Raf-1 substrate in vitro and did not significantly affect Raf-1 kinase activity in a purified system. However, in cell-free extracts 14-3-3 acted as a Ras-independent activator of both c-Raf-1 and the Raf-1 kinase domain. The same results were obtained in vivo using transfection assays; 14-3-3 enhanced both c-Raf-1- and Raf-1 kinase domain-stimulated expression of AP-1- and NF-kappa B-dependent reporter genes and accelerated Raf-1 kinase domain-triggered differentiation of PC12 cells. We conclude that 14-3-3 is a latent co-activator bound to unactivated Raf-1 in quiescent cells and mediates mitogen-triggered but Ras-independent regulatory effects aimed directly at the kinase domain. 相似文献
96.
Caccone A; Moriyama EN; Gleason JM; Nigro L; Powell JR 《Molecular biology and evolution》1996,13(9):1224-1232
Drosophila melanogaster belongs to a closely related group of eight species
collectively known as the melanogaster subgroup; all are native to
sub-Saharan Africa and islands off the east coast of Africa. The
phylogenetic relationships of most species in this subgroup have been well
documented; however, the three most closely related species, D. simulans,
D. sechellia, and D. mauritiana, have remained problematic from a
phylogenetic standpoint as no data set has unambiguously resolved them. We
present new DNA sequence data on the nullo and Serendipity-alpha genes and
combine them with all available nuclear DNA sequence data; the total data
encompass 12 genes and the ITS of rDNA. A methodological problem arose
because nine of the genes had information on intraspecific polymorphisms in
at least one species. We explored the effect of inclusion/exclusion of
polymorphic sites and found that it had very little effect on phylogenetic
inferences, due largely to the fact that 82% of polymorphisms are
autapomorphies (unique to one species). We have also reanalyzed our
previous DNA-DNA hybridization data with a bootstrap procedure. The
combined sequence data set and the DNA-DNA hybridization data strongly
support the sister status of the two island species, D. sechellia and D.
mauritiana. This at least partially resolves what had been a paradox of
parallel evolution in these two species.
相似文献
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INTRoDUCTIoNlYho1iumrePensL,whiteclover,isaneconomicallyimportantplantspeciesintemperatepastures.Asbrieflyreportedby[1],ithas16pairsofchromosomes(2n=32).Asyet,nodetailedcytologicalexaminationofthisspecies,suchasC-banding,hasbeenrep0rted.Inthelastdecade,thetechnique0fC-bandinghasbeenusedt0examinehighlyrepeatedsequencesinplantchrom0s0mesandhasprovidedausefultoolf0rtheanalysis0fcyt0geneticstructureincr0pplants[2-71.Inplants,thechr0m0s0mall0calizationofhighlyrepeatedDNAsequencesbyinsituhybr… 相似文献
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The period (per) locus has received much attention in molecular evolution
studies because it is one of the best studied "behavioral genes" and
because it offers insight into the evolution of repetitive sequences. We
studied most of the coding region of per in Drosophila willistoni and
confirmed previously observed patterns of conservation and divergence among
distantly related species. Five regions are so highly diverged that they
cannot be aligned, whereas a region encompassing the PAS domain is very
conserved. Structural and nucleotide polymorphism patterns in the
willistoni group are not the same as those observed in previously studied
species. We sequenced the region homologous to the highly polymorphic
threonine-glycine repeat of D. melanogaster in multiple strains of D.
willistoni, as well as in other members of willistoni group, and found an
unusual amount of conservation in this region. However, the next
nonconserved region downstream in the sequence is quite variable and
polymorphic for the number of repeated glycines. The glycine codon usage is
significantly different in this glycine repeat as compared to other parts
of the gene. We were able to plot the directionality of change in the
glycine repeat region onto a phylogeny and find that the addition of
glycines is the general trend with the diversification of the willistoni
group.
相似文献