Structural and functional interactions between measles virus nucleocapsid protein and cellular heat shock protein

Structural and functional interactions between measles virus nucleocapsid protein and cellular heat shock protein

Bibliographic Details
Main Author: Zhang, Xinsheng
Language:English
Published: The Ohio State University / OhioLINK 2004
Subjects:
Hsp72
Morbillivirus
Nucleocapsid protein
Binding motif
Reverse genetics
Biacore
Online Access:http://rave.ohiolink.edu/etdc/view?acc_num=osu1078417800
id ndltd-OhioLink-oai-etd.ohiolink.edu-osu1078417800
record_format oai_dc
spelling ndltd-OhioLink-oai-etd.ohiolink.edu-osu10784178002021-08-03T05:48:42Z Structural and functional interactions between measles virus nucleocapsid protein and cellular heat shock protein Zhang, Xinsheng Hsp72 Morbillivirus Nucleocapsid protein Binding motif Reverse genetics Biacore Cells respond to stress by producing heat shock proteins (HSPs), particularly the major inducible 72kDa protein (Hsp72). Although HSPs are generally viewed as protecting cells against injury, data has also shown that elevated cellular Hsp72 promotes damage induced by measles virus (MV) in vitro. Specifically, Hsp72 enhances MV transcription and replication activity, resulting in increased viral antigen expression, cytopathic effect, and infectious viral progeny release. Hsp72-dependent stimulation of MV transcription reflects functional interactions between Hsp72 and an 8 amino acid (aa) binding motif within the extreme C-terminus of the nucleocapsid protein (N), a major structural component of the nucleocapsid. Nucleocapsid consists of viral genomic RNA packaged by N, polymerase cofactor (P), and the viral encoded RNA-dependent RNA polymerase (L). The nucleocapsid activity can be measured using minireplicons in which the viral genomic coding sequence is replaced by chloramphenical acetyltransferase (CAT) RNA. Using this approach, we identified the C-terminal 24 amino acids of N protein as a negative regulatory domain of MV transcription. Addition of Hsp72 by transfection significantly stimulates viral transcription/replication activity mediated by N. Mutations in the Hsp72 binding motif based upon naturally occurring sequence polymorphisms can diminish Hsp72 stimulation of MV minireplicon reporter gene expression. Loss of functional interaction correlates to the loss of low affinity binding interaction. Binding interactions were monitored in real time using surface plasmon resonance technology (BIAcore). These same mutations, when incorporated into the N protein of recombinant infectious virus, also abrogate Hsp72-dependent infection phenotypes. Incorporation of a non-functional Hsp72 binding motif reduced the stimulatory effect of pre-conditioning upon viral transcription, cytopathic effect (CPE), and cell-free infectious viral progeny release. However, loss of a functional binding motif doesn’t abrogate the stimulatory effect of pre-conditioning on viral genome levels. N protein lacking the Hsp72 binding motif maintains the ability to form a stable complex with Hsp72 by both nucleocapsid-Hsp72 particle isolation as well as N-Hsp72 co-immunoprecipitation. BIAcore analysis of the last 125 aa of N protein (Ntail) and Hsp72 interaction confirmed the existence of additional Hsp72 binding domains. The relevance of high affinity interactions to Hsp72-dependent stimulation of MV genome replication remains to be established. 2004-03-09 English text The Ohio State University / OhioLINK http://rave.ohiolink.edu/etdc/view?acc_num=osu1078417800 http://rave.ohiolink.edu/etdc/view?acc_num=osu1078417800 unrestricted This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws.
collection NDLTD
language English
sources NDLTD
topic Hsp72
Morbillivirus
Nucleocapsid protein
Binding motif
Reverse genetics
Biacore
spellingShingle Hsp72
Morbillivirus
Nucleocapsid protein
Binding motif
Reverse genetics
Biacore
Zhang, Xinsheng
Structural and functional interactions between measles virus nucleocapsid protein and cellular heat shock protein
author Zhang, Xinsheng
author_facet Zhang, Xinsheng
author_sort Zhang, Xinsheng
title Structural and functional interactions between measles virus nucleocapsid protein and cellular heat shock protein
title_short Structural and functional interactions between measles virus nucleocapsid protein and cellular heat shock protein
title_full Structural and functional interactions between measles virus nucleocapsid protein and cellular heat shock protein
title_fullStr Structural and functional interactions between measles virus nucleocapsid protein and cellular heat shock protein
title_full_unstemmed Structural and functional interactions between measles virus nucleocapsid protein and cellular heat shock protein
title_sort structural and functional interactions between measles virus nucleocapsid protein and cellular heat shock protein
publisher The Ohio State University / OhioLINK
publishDate 2004
url http://rave.ohiolink.edu/etdc/view?acc_num=osu1078417800
work_keys_str_mv AT zhangxinsheng structuralandfunctionalinteractionsbetweenmeaslesvirusnucleocapsidproteinandcellularheatshockprotein
_version_ 1719425921873608704

Similar Items