Systematic functional analysis of Bicaudal-D serine phosphorylation and intragenic suppression of a female sterile allele of BicD.
Protein phosphorylation is involved in posttranslational control of essentially all biological processes. Using mass spectrometry, recent analyses of whole phosphoproteomes led to the identification of numerous new phosphorylation sites. However, the function of most of these sites remained unknown. We chose the Drosophila Bicaudal-D protein to estimate the importance of individual phosphorylation events. Being involved in different cellular processes, BicD is required for oocyte determination, for RNA transport during oogenesis and embryogenesis, and for photoreceptor nuclei migration in the developing eye. The numerous roles of BicD and the available evidence for functional importance of BicD phosphorylation led us to identify eight phosphorylation sites of BicD, and we tested a total of 14 identified and suspected phosphoserine residues for their functional importance in vivo in flies. Surprisingly, all these serines turned out to be dispensable for providing sufficient basal BicD activity for normal growth and development. However, in a genetically sensitized background where the BicD(A40V) protein variant provides only partial activity, serine 103 substitutions are not neutral anymore, but show surprising differences. The S103D substitution completely inactivates the protein, whereas S103A behaves neutral, and the S103F substitution, isolated in a genetic screen, restores BicD(A40V) function. Our results suggest that many BicD phosphorylation events may either be fortuitous or play a modulating function as shown for Ser(103). Remarkably, amongst the Drosophila serines we found phosphorylated, Ser(103) is the only one that is fully conserved in mammalian BicD.
Koch
Rafael
R
Institute of Cell Biology, University of Bern, Bern, Switzerland.
Ledermann
Romana
R
Urwyler
Olivier
O
Heller
Manfred
M
Suter
Beat
B
D000483:Alleles; D000818:Animals; D001665:Binding Sites; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D047108:Embryonic Development; D005260:Female; D020125:Mutation, Missense; D009865:Oocytes; D009866:Oogenesis; D010766:Phosphorylation / Q000502:physiology; D010768:Phosphoserine; D012694:Serine / Q000378:metabolism
C061998:BicD protein, Drosophila; D029721:Drosophila Proteins; D010768:Phosphoserine; D012694:Serine