Characterization of the microtubule proteome during post-diapause development of Artemia franciscana.
The microtubule proteome encompasses tubulin and a diverse group of proteins which associate with tubulin upon microtubule formation. These proteins either determine microtubule organization and function or their activity is influenced by microtubule association. To characterize the microtubule proteome in Artemia franciscana, tubulin assembly was induced with taxol in vitro after 0 and 12 h of post-diapause development. Proteins obtained by extraction of microtubules with 0.5 M NaCl were electrophoresed in two-dimensional gels and analyzed by mass spectrometry. Fifty-five proteins were identified with 10 of these occurring at both developmental stages, and multiple isoforms were observed for some proteins of the Artemia proteome. Their functions include roles in membrane transport, metabolism, chaperoning and protein synthesis, thus reflecting physiological properties of encysted Artemia such as stress resistance and the ability to rapidly initiate post-diapause development. For example, chaperones may protect tubulin during encystment and facilitate folding in metabolically active embryos. Additionally, the interaction of metabolic enzymes with microtubules funnels reaction intermediates, potentially enhancing efficiency within biochemical processes. This study represents the first systematic characterization of a crustacean microtubule proteome. Although it is difficult to be certain that all protein associations documented herein occur in vivo, the results suggest how protein-protein interactions contribute to cytoplasmic organization while implying how Artemia embryos resist stress and remain capable of development once diapause terminates.
O'Connell
Paul A
PA
Department of Biology, Dalhousie University, Halifax, Nova Scotia, Canada B3H 4J1.
Pinto
Devanand M
DM
Chisholm
Ken A
KA
MacRae
Thomas H
TH
D000818:Animals; D001156:Artemia / Q000737:chemistry / Q000196:embryology* / Q000378:metabolism*; D008870:Microtubules / Q000737:chemistry* / Q000648:ultrastructure; D025941:Protein Interaction Mapping; D020543:Proteome / Q000737:chemistry* / Q000378:metabolism*; D040901:Proteomics / Q000379:methods; D014404:Tubulin / Q000737:chemistry / Q000378:metabolism
D020543:Proteome; D014404:Tubulin