D-raf, Draf, phl, pole hole, l(1)ph
serine/threonine-protein kinase - an effector of Ras - directs the RAF/MEK/ERK pathway to regulate cell proliferation, differentiation and survival downstream of receptor tyrosine kinases such as Torso, Epidermal growth factor receptor, and Sevenless.
Gene model reviewed during 5.51
Gene model reviewed during 5.46
Gene model reviewed during 5.53
Interacts with Dsor1/MEK1 and ksr; Dsor1 binding to ksr probably promotes ksr and Raf dimerization and ksr-mediated Raf transactivation.
Extensively phosphorylated 1 to 2 hours after egg laying.
Click to get a list of regulatory features (enhancers, TFBS, etc.) and gene disruptions (point mutations, indels, etc.) within or overlapping Dmel\Raf using the Feature Mapper tool.
GBrowse - Visual display of RNA-Seq signalsView Dmel\Raf in GBrowse 2
Please Note FlyBase no longer curates genomic clone accessions so this list may not be complete
Please Note This section lists cDNAs and ESTs that fall within the genomic extent of the gene model, which may include cDNAs and ESTs of genes within introns, or of overlapping genes. Please see GBrowse for alignment of the cDNAs and ESTs to the gene model.
For each fully sequenced cDNA the DGRC maintains various forms of the cDNA (e.g tagged or untagged) in several different host vectors for subsequent cloning and expression in Drosophila and Drosophila cell lines.
dsRNA made from templates generated with primers directed against this gene.
dsRNA made from templates generated with primers directed against this gene tested in RNAi screen for effects on Kc167 and S2R+ cell morphology.
A basal level of phl activity is required for the accumulation of circulating blood cells, while high level activity of phl is required for the generation of differentiated lamellocytes in larvae. phl acts downstream of hop during differentiation of lamellocytes in larvae.
The MAPK cascade is required for Ras85D mitogenic response, loss of function mutations in phl, Dsor1, rl and ksr dominantly suppress hyperplastic growth, as do mutations in the Ras85D effector loop that disrupt the Ras85D-phl interaction.
In a sample of 79 genes with multiple introns, 33 showed significant heterogeneity in G+C content among introns of the same gene and significant positive correspondence between the intron and the third codon position G+C content within genes. These results are consistent with selection adding against preferred codons at the start of genes.
The role of Ras85D in phl activation is not limited to the translocation of phl to the membrane through a Ras85D-phl association. Ras85D is essential for the activation of an additional factor which in turn activates phl.
Genetic studies suggest that 14-3-3ε functions in multiple receptor tyrosine kinase pathways, acting downstream or parallel to phl, but upstream of aop and phyl, two nuclear factors involved in Ras85D signalling.
Endogenous mutant forms of phl may compete with wild-type phl for a positive factor required for its activation and/or membrane association or block access to it substrate(s) thereby affecting rescue of the tor pathway.
Two sequences homologous to DRE (DNA replication-related element) in the 5' flanking region of the phl gene are identified and transcriptional activity is confirmed through gel mobility shift assays, transient Ecol\CAT assays and spatial expression of Ecol\lacZ in transgenic larval tissues. phl is another target of zen protein in cultured cells.
In vitro studies reveal amino acid S743 is essential for phl function in embryos. Altered forms of phl have distinct activity profiles indicating that each structural modification differentially affects the regulation and/or propagation of the tor signal by the mutant phl proteins.
The membrane targetted phl kinase generates a signal that directs terminal development.
Scer\GAL4-UAS system has been used to demonstrate that activating phl in the follicle cells is sufficient to dorsalise the egg suggesting that phl is required in the follicle cells, rather than in the oocyte, to specify dorsoventral polarity. Mutant combinations of phl and Egfr demonstrates that phl acts downstream of Egfr.
Clonal analysis shows that mutations that alter conserved residues in the phl product known to be necessary for kinase activity are associated with a null phenotype, demonstrating that phl activity is necessary for its role in torso signalling. Weaker alleles show phenotype similar to corkscrew maternal effect phenotype. Two mutations affecting late phl functions imply a novel role for phl in cell fate establishment in the eye.
Germline clone analysis with an artificial bcd responder gene (three bcd consensus binding sites driving Ecol\lacZ) indicates that the bcd responder is not efficiently repressed in phl mutants, and that phl may be involved in the kinase pathway mediating inhibition of bcd-dependent activation.
Biochemical analysis of the signal transduction pathway determining terminal structure development.
The phl serine/threonine kinase plays a crucial role in the R7 pathway: the response to sev gene product activity is dependent on phl function, and a constitutively activated phl protein can induce R7 cell development in the absence of sev function. Genetic evidence suggests that phl acts downstream of Ras85D and upstream of sina in this signal transduction pathway.
csw and phl acts in concert to regulate tll expression.
The maternal terminal system is necessary to activate tll expression in the terminal caps.
Comparison of CpG distribution in the coding region of 121 genes from six species supports the mCpG mutational hotspot explanation of CpG suppression in methylated species at position II-III and III-I.
phl is zygotically expressed and encodes a potential Ser/Thr kinase. phl acts downstream of tor.
phl is essential for continued cell proliferation of diploid cells in the larva.
The wild-type allele of phl seems to be involved in the setting up of positional values in embryos and also in the proliferation of diploid cells in imaginal discs (Ambrosio, Mahowald and Perrimon, 1989a). phl mutants are recessive early-pupal lethals, which display very small imaginal discs. Embryos derived from germ-line clones and lacking phl+ activity show the 'torso' or 'pole hole' phenotype; structures at the anterior and the posterior end (spiracles, anal tufts and the entire eighth abdominal segment) fail to develop (Perrimon, Engstrom and Mahowald, 1984a; Perrimon, Engstrom and Mahowald, 1984b; Ambrosio, Mahowald and Perrimon, 1989a). The fate map of the blastoderm is shifted posteriorly and fewer segments with more cells result. A partial rescue of these mutants has been obtained with phl+ sperm (Ambrosio, Engstrom and Mahowald); all structures posterior to abdominal segment seven are missing (Perrimon and Mahowald, 1986).