Document Type
Article
Language
eng
Format of Original
9 p.
Publication Date
11-15-2010
Publisher
Elsevier
Source Publication
Developmental Biology
Source ISSN
0012-1606
Abstract
The vitelline membrane (VM), the oocyte proximal layer of the Drosophila eggshell, contains four major proteins (VMPs) that possess a highly conserved “VM domain” which includes three precisely spaced, evolutionarily conserved, cysteines (CX7CX8C). Focusing on sV23, this study showed that the three cysteines are not functionally equivalent. While substitution mutations at the first (C123S) or third (C140S) cysteines were tolerated, females with a substitution at the second position (C131S) were sterile. Fractionation studies showed that sV23 incorporates into a large disulfide linked network well after its secretion ceases, suggesting that post-depositional mechanisms are in place to restrict disulfide bond formation until late oogenesis, when the oocyte no longer experiences large volume increases. Affinity chromatography utilizing histidine tagged sV23 alleles revealed small sV23 disulfide linked complexes during the early stages of eggshell formation that included other VMPs, namely sV17 and Vml. The early presence but late loss of these associations in an sV23 double cysteine mutant suggests that reorganization of disulfide bonds may underlie the regulated growth of disulfide linked networks in the vitelline membrane. Found within the context of a putative thioredoxin active site (CXXS) C131, the critical cysteine in sV23, may play an important enzymatic role in isomerizing intermolecular disulfide bonds during eggshell assembly.
Recommended Citation
Wu, T.; Manogaran, Anita L.; Beauchamp, J. M.; and Waring, Gail L., "Drosophila Vitelline Membrane Assembly: A Critical Role for an Evolutionarily Conserved Cysteine in the “VM domain” of sV23" (2010). Biological Sciences Faculty Research and Publications. 296.
https://epublications.marquette.edu/bio_fac/296
Comments
Accepted version. Developmental Biology, Vol. 347, No. 2 (November 15, 2010): 360-368. DOI. © 2010 Elsevier. Used with permission.
NOTICE: this is the author’s version of a work that was accepted for publication in Developmental Biology. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Developmental Biology, VOL 347, ISSUE 2, November 2010, DOI.