Characterization of a calcium binding domain in the VP1 protein of the avian polyomavirus, budgerigar fledgling disease virus.
Calcium ions appear to play a major role in maintaining the structural integrity and assembly of papovavirus virions and are likely involved in the process of viral uncoating. Recently it was reported that the purified recombinant VP1 protein of budgerigar fledgling disease virus (BFDV) was capable of assembling into capsid-like particles in the presence of calcium. It is now reported that the major capsid protein VP1 of BFDV binds calcium ions in an in vitro calcium binding assay. Two deletions were made in the VP1 protein to identify a calcium binding domain and to further characterize the role of calcium ions in the capsid assembly process. Recombinant VP1 lacking a putative calcium binding domain (Asp-237-Asp-248) failed to bind radioactive 45Ca2+ yet associated into capsomeres. These capsomeres were similar in shape to the wild-type VP1 but were unable to assemble into capsid-like particles. Likewise, recombinant VP1 lacking ten carboxyl terminal amino acids (Glu-334-Arg-343) also formed capsomeres that were unable to assemble into capsid-like particles. In contrast to the VP1 protein with the internal deletion, the protein with the truncated carboxyl terminus bound 45Ca2+ in the in vitro assay. These results have identified a calcium binding domain (Asp-237-Asp-248) for the BFDV VP1 protein and a crucial role for the VP1 carboxyl terminal amino acids (Glu-334-Arg-343) in capsid assembly.
Rodgers
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Division of Biology, Kansas State University, Manhattan 66506, USA.
Consigli
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D000818:Animals; D001665:Binding Sites; D001717:Birds / Q000821:virology; D002118:Calcium / Q000378:metabolism*; D002213:Capsid / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D036022:Capsid Proteins; D002642:Chick Embryo; D011120:Polyomavirus / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D014771:Virion / Q000378:metabolism
D036022:Capsid Proteins; D011993:Recombinant Fusion Proteins; C062481:VP1 protein, polyomavirus; D002118:Calcium