Loss of Dictyostelium HSPC300 causes a scar-like phenotype and loss of SCAR protein.
BACKGROUND
SCAR/WAVE proteins couple signalling to actin polymerization, and are thus fundamental to the formation of pseudopods and lamellipods. They are controlled as part of a five-membered complex that includes the tiny HSPC300 protein. It is not known why SCAR/WAVE is found in such a large assembly, but in Dictyostelium the four larger subunits have different, clearly delineated functions.


RESULTS
We have generated Dictyostelium mutants in which the HSPC300 gene is disrupted. As has been seen in other regulatory complex mutants, SCAR is lost in these cells, apparently by a post-translational mechanism, though PIR121 levels do not change. HSPC300 knockouts resemble scar mutants in slow migration, roundness, and lack of large pseudopods. However hspc300-colonies on bacteria are larger and more similar to wild type, suggesting that some SCAR function can survive without HSPC300. We find no evidence for functions of HSPC300 outside the SCAR complex.


CONCLUSIONS
HSPC300 is essential for most SCAR complex functions. The phenotype of HSPC300 knockouts is most similar to mutants in scar, not the other members of the SCAR complex, suggesting that HSPC300 acts most directly on SCAR itself.
Pollitt
Alice Y
AY
School of Biosciences, University of Birmingham, Birmingham, B15 2TT, UK. a.y.pollitt@bham.ac.uk
Insall
Robert H
RH
D000595:Amino Acid Sequence; D000818:Animals; D004023:Dictyostelium / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D017125:Genes, Protozoan; D008969:Molecular Sequence Data; D009068:Movement; D046912:Multiprotein Complexes; D009154:Mutation; D010641:Phenotype; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction
D046912:Multiprotein Complexes; D015800:Protozoan Proteins; C491975:SCAR protein, Dictyostelium