A mature and fusogenic form of the Nipah virus fusion protein requires proteolytic processing by cathepsin L

Cara Theresia Pager; Willie Warren Craft; Jared Patch; Rebecca Ellis Dutch

doi:10.1016/j.virol.2006.01.007

A mature and fusogenic form of the Nipah virus fusion protein requires proteolytic processing by cathepsin L

Cara Theresia Pager, Willie Warren Craft, Jared Patch, Rebecca Ellis Dutch

Research output: Contribution to journal › Article › peer-review

131 Scopus citations

Abstract

The Nipah virus fusion (F) protein is proteolytically processed to F ₁ + F₂ subunits. We demonstrate here that cathepsin L is involved in this important maturation event. Cathepsin inhibitors ablated cleavage of Nipah F. Proteolytic processing of Nipah F and fusion activity was dramatically reduced in cathepsin L shRNA-expressing Vero cells. Additionally, Nipah virus F-mediated fusion was inhibited in cathepsin L-deficient cells, but coexpression of cathepsin L restored fusion activity. Both purified cathepsin L and B could cleave immunopurified Nipah F protein, but only cathepsin L produced products of the correct size. Our results suggest that endosomal cathepsins can cleave Nipah F, but that cathepsin L specifically converts Nipah F to a mature and fusogenic form.

Original language	English
Pages (from-to)	251-257
Number of pages	7
Journal	Virology
Volume	346
Issue number	2
DOIs	https://doi.org/10.1016/j.virol.2006.01.007
State	Published - Mar 15 2006

Bibliographical note

Funding Information:
We would like to thank Dr. Lin-fa Wang (Australian Animal Health Laboratory) for the Nipah F and G plasmids; Drs. Paul Selleck and Chris Morrissy for the anti-gamma inactivated Nipah virus antiserum (Australian Animal Health Laboratory); Dr. Doug Andres (University of Kentucky) for the pSuper-Scramble plasmid; Karl-Klaus Conzelman (Pettenkofer Institut) for the BSR cells; and Dr. Terence Dermody (Vanderbilt University) for the cathepsin L MEFs and pSG5-cathepsin L vector. We greatly appreciate assistance from Jennifer Strange in the UK Flow Cytometry Core Facility. We are grateful to members of the Dutch lab and Dr. Christopher Broder for critically reviewing the manuscript. C.T.P. and W.W.C. are recipients of an AHA Ohio Valley Affiliate predoctoral fellowship and a Ruth L. Kirshechstein National Research Service Award, respectively. This study was supported by NIAID grant AI063052 to R.E.D.

Funding

We would like to thank Dr. Lin-fa Wang (Australian Animal Health Laboratory) for the Nipah F and G plasmids; Drs. Paul Selleck and Chris Morrissy for the anti-gamma inactivated Nipah virus antiserum (Australian Animal Health Laboratory); Dr. Doug Andres (University of Kentucky) for the pSuper-Scramble plasmid; Karl-Klaus Conzelman (Pettenkofer Institut) for the BSR cells; and Dr. Terence Dermody (Vanderbilt University) for the cathepsin L MEFs and pSG5-cathepsin L vector. We greatly appreciate assistance from Jennifer Strange in the UK Flow Cytometry Core Facility. We are grateful to members of the Dutch lab and Dr. Christopher Broder for critically reviewing the manuscript. C.T.P. and W.W.C. are recipients of an AHA Ohio Valley Affiliate predoctoral fellowship and a Ruth L. Kirshechstein National Research Service Award, respectively. This study was supported by NIAID grant AI063052 to R.E.D.

Funders	Funder number
American the American Heart Association
National Institute of Allergy and Infectious Diseases	R21AI063052, R01AI051517
Not added	12720

Keywords

Cathepsin L
Fusion protein
Nipah
Proteolytic processing

ASJC Scopus subject areas

Virology

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10.1016/j.virol.2006.01.007

Cite this

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title = "A mature and fusogenic form of the Nipah virus fusion protein requires proteolytic processing by cathepsin L",

abstract = "The Nipah virus fusion (F) protein is proteolytically processed to F 1 + F2 subunits. We demonstrate here that cathepsin L is involved in this important maturation event. Cathepsin inhibitors ablated cleavage of Nipah F. Proteolytic processing of Nipah F and fusion activity was dramatically reduced in cathepsin L shRNA-expressing Vero cells. Additionally, Nipah virus F-mediated fusion was inhibited in cathepsin L-deficient cells, but coexpression of cathepsin L restored fusion activity. Both purified cathepsin L and B could cleave immunopurified Nipah F protein, but only cathepsin L produced products of the correct size. Our results suggest that endosomal cathepsins can cleave Nipah F, but that cathepsin L specifically converts Nipah F to a mature and fusogenic form.",

keywords = "Cathepsin L, Fusion protein, Nipah, Proteolytic processing",

author = "Pager, \{Cara Theresia\} and Craft, \{Willie Warren\} and Jared Patch and Dutch, \{Rebecca Ellis\}",

note = "Funding Information: We would like to thank Dr. Lin-fa Wang (Australian Animal Health Laboratory) for the Nipah F and G plasmids; Drs. Paul Selleck and Chris Morrissy for the anti-gamma inactivated Nipah virus antiserum (Australian Animal Health Laboratory); Dr. Doug Andres (University of Kentucky) for the pSuper-Scramble plasmid; Karl-Klaus Conzelman (Pettenkofer Institut) for the BSR cells; and Dr. Terence Dermody (Vanderbilt University) for the cathepsin L MEFs and pSG5-cathepsin L vector. We greatly appreciate assistance from Jennifer Strange in the UK Flow Cytometry Core Facility. We are grateful to members of the Dutch lab and Dr. Christopher Broder for critically reviewing the manuscript. C.T.P. and W.W.C. are recipients of an AHA Ohio Valley Affiliate predoctoral fellowship and a Ruth L. Kirshechstein National Research Service Award, respectively. This study was supported by NIAID grant AI063052 to R.E.D.",

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AU - Pager, Cara Theresia

AU - Craft, Willie Warren

AU - Patch, Jared

AU - Dutch, Rebecca Ellis

N1 - Funding Information: We would like to thank Dr. Lin-fa Wang (Australian Animal Health Laboratory) for the Nipah F and G plasmids; Drs. Paul Selleck and Chris Morrissy for the anti-gamma inactivated Nipah virus antiserum (Australian Animal Health Laboratory); Dr. Doug Andres (University of Kentucky) for the pSuper-Scramble plasmid; Karl-Klaus Conzelman (Pettenkofer Institut) for the BSR cells; and Dr. Terence Dermody (Vanderbilt University) for the cathepsin L MEFs and pSG5-cathepsin L vector. We greatly appreciate assistance from Jennifer Strange in the UK Flow Cytometry Core Facility. We are grateful to members of the Dutch lab and Dr. Christopher Broder for critically reviewing the manuscript. C.T.P. and W.W.C. are recipients of an AHA Ohio Valley Affiliate predoctoral fellowship and a Ruth L. Kirshechstein National Research Service Award, respectively. This study was supported by NIAID grant AI063052 to R.E.D.

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N2 - The Nipah virus fusion (F) protein is proteolytically processed to F 1 + F2 subunits. We demonstrate here that cathepsin L is involved in this important maturation event. Cathepsin inhibitors ablated cleavage of Nipah F. Proteolytic processing of Nipah F and fusion activity was dramatically reduced in cathepsin L shRNA-expressing Vero cells. Additionally, Nipah virus F-mediated fusion was inhibited in cathepsin L-deficient cells, but coexpression of cathepsin L restored fusion activity. Both purified cathepsin L and B could cleave immunopurified Nipah F protein, but only cathepsin L produced products of the correct size. Our results suggest that endosomal cathepsins can cleave Nipah F, but that cathepsin L specifically converts Nipah F to a mature and fusogenic form.

AB - The Nipah virus fusion (F) protein is proteolytically processed to F 1 + F2 subunits. We demonstrate here that cathepsin L is involved in this important maturation event. Cathepsin inhibitors ablated cleavage of Nipah F. Proteolytic processing of Nipah F and fusion activity was dramatically reduced in cathepsin L shRNA-expressing Vero cells. Additionally, Nipah virus F-mediated fusion was inhibited in cathepsin L-deficient cells, but coexpression of cathepsin L restored fusion activity. Both purified cathepsin L and B could cleave immunopurified Nipah F protein, but only cathepsin L produced products of the correct size. Our results suggest that endosomal cathepsins can cleave Nipah F, but that cathepsin L specifically converts Nipah F to a mature and fusogenic form.

KW - Cathepsin L

KW - Fusion protein

KW - Nipah

KW - Proteolytic processing

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A mature and fusogenic form of the Nipah virus fusion protein requires proteolytic processing by cathepsin L

Abstract

Bibliographical note

Funding

Keywords

ASJC Scopus subject areas

Access to Document

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