Subunit cleavage of mosquito pro-vitellogenin by a subtilisin-like convertase.
The eukaryotic convertase family plays an important role in posttranslational proteolytic processing and activation of many pro- and polypeptides that have at their cleavage sites the paired basic motif, RX(K/R)R. Recent studies have revealed that the cleavage site of insect pro-vitellogenins (pro-Vg) also contains this motif. To identify and characterize the insect pro-Vg processing enzyme, Vg convertase (VC), its cDNA was cloned from a vitellogenic female fat body cDNA library of the mosquito, Aedes aegypti. The 3735-bp-long VC cDNA has an open reading frame encoding a 115-kDa protein. In vitro transcription/translation of VC cDNA revealed that this 115-kDa protein becomes 140 kDa after co- and posttranslational modifications. The VC deduced amino acid sequence has high similarity to and a domain structure characteristic of furin-like convertases. Northern blot analysis showed that a single 4.2-kb transcript was expressed in the fat body during the first 18 hr of the Vg synthetic period. Coexpression of VC cDNA with mosquito Vg cDNA resulted in correct cleavage of pro-Vg. Thus, this newly identified convertase is, indeed, a functional fat body-specific enzyme for pro-Vg cleavage.
Chen
J S
JS
Program in Genetics and Department of Entomology, Michigan State University, East Lansing, 48824-1115, USA.
Raikhel
A S
AS
D000595:Amino Acid Sequence; D000818:Animals; D000852:Anopheles / Q000378:metabolism*; D001483:Base Sequence; D018076:DNA, Complementary; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism*; D005216:Fat Body / Q000201:enzymology; D005260:Female; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D011498:Protein Precursors / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D013379:Substrate Specificity; D013381:Subtilisins / Q000378:metabolism*; D014158:Transcription, Genetic; D014819:Vitellogenins / Q000378:metabolism*
D018076:DNA, Complementary; D011498:Protein Precursors; D012333:RNA, Messenger; D014819:Vitellogenins; D010450:Endopeptidases; D013381:Subtilisins; C099719:Vg convertase