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Processing of nonstructural proteins NS4A and NS4B of dengue 2 virus in vitro and in vivo

Preugschat, Frank and Strauss, James H. (1991) Processing of nonstructural proteins NS4A and NS4B of dengue 2 virus in vitro and in vivo. Virology, 185 (2). pp. 689-697. ISSN 0042-6822. https://resolver.caltech.edu/CaltechAUTHORS:20160203-164500016

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Abstract

The production, from polyprotein precursors, of two hydrophobic nonstructural proteins of dengue 2 (DEN2) virus, NS4A and NS4B, was analyzed both in cell-free systems and in infected cells. In DEN2-infected cells, NS4B is first produced as a peptide of apparent size 30 kDa; NS4B is then post-translationally modified, in an unknown way, to produce a polypeptide of apparent size 28 kDa. The rate and extent of NS4B modification was found to be cell-dependent; in BHK cells the half-time for the conversion of the 30-kDa form to the 28-kDa form was 90 min. N-terminal sequence analysis of NS4B suggests that the N-terminus is produced by an enzyme with a specificity similar to that of signalase. Low levels of a putative polyprotein, NS4AB, were also found in mammalian cells, but not mosquito cells, infected with DEN2, suggesting that a small proportion of DEN2 4A/4B cleavage can occur post-translationally or that some nonstructural polyproteins escape normal processing. Cleavage of the 4A/4B bond in infected cells required expression of DEN2 sequences in addition to those in NS4A and NS4B, as NS4AB produced in cells by a vaccinia expression system was not cleaved. NS4AB produced in cells by a vaccinia expression system was modified post-translationally, presumably in the same way as NS4B. We show that upon translation of DEN2 polyproteins in a cell-free system, the N-terminus of NS4A is generated by cleavage by the viral nonstructural proteinase NS3 and that processing of DEN2 polyproteins occurs with a preferred, but nonobligatory order.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1016/0042-6822(91)90540-RDOIArticle
http://www.sciencedirect.com/science/article/pii/004268229190540RPublisherArticle
Additional Information:© 1991 Academic Press, Inc. Received July 16, 1991; accepted August 21, 1991. The authors thank Ellen Strauss. Richard Kuhn, and Ron Weir for stimulating discussions and editorial expertise; David Teplow and Tammy Bauer for their expertise and assistance with protein sequencing; and Joel Dalrymple for furnishing cells, virus, and hyperimmune antiserum. This work was supported by Grant V22/181/10 from the World Health Organization and by Grant AI20612 from the National Institutes of Health.
Funders:
Funding AgencyGrant Number
World Health OrganizationV22/181/10
NIHAI20612
Issue or Number:2
Record Number:CaltechAUTHORS:20160203-164500016
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20160203-164500016
Official Citation:Frank Preugschat, James H. Strauss, Processing of nonstructural proteins NS4A and NS4B of dengue 2 virus in vitro and in vivo, Virology, Volume 185, Issue 2, 1991, Pages 689-697, ISSN 0042-6822, http://dx.doi.org/10.1016/0042-6822(91)90540-R. (http://www.sciencedirect.com/science/article/pii/004268229190540R)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:64218
Collection:CaltechAUTHORS
Deposited By: Donna Wrublewski
Deposited On:04 Feb 2016 01:07
Last Modified:03 Oct 2019 09:35

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