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PMID: 12768027 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S. Retracted Publication

Basic residues of the helix six domain of influenza virus M1 involved in nuclear translocation of M1 can be replaced by PTAP and YPDL late assembly domain motifs.

Journal of virology ·Vol. 77 ·No. 12 ·2003-06-00 ·Pages 7078-92

Hui EK, Barman S, Yang TY, Nayak DP

Abstract

Influenza type A virus matrix (M1) protein possesses multiple functional motifs in the helix 6 (H6) domain (amino acids 91 to 105), including nuclear localization signal (NLS) (101-RKLKR-105) involved in translocating M1 from the cytoplasm into the nucleus. To determine the role of the NLS motif in the influenza virus life cycle, we mutated these and the neighboring sequences by site-directed mutagenesis, and influenza virus mutants were generated by reverse genetics. Our results show that infectious viruses were rescued by reverse genetics from all single alanine mutations of amino acids in the H6 domain and the neighboring region except in three positions (K104A and R105A within the NLS motif and E106A in loop 6 outside the NLS motif). Among the rescued mutant viruses, R101A and R105K exhibited reduced growth and small-plaque morphology, and all other mutant viruses showed the wild-type phenotype. On the other hand, three single mutations (K104A, K105A, and E106A) and three double mutations (R101A/K102A, K104A/K105A, and K102A/R105A) failed to generate infectious virus. Deletion (Delta YRKL) or mutation (4A) of YRKL also abolished generation of infectious virus. However, replacement of the YRKL motif with PTAP or YPDL as well as insertion of PTAP after 4A mutation yielded infectious viruses with the wild-type phenotype. Furthermore, mutant M1 proteins (R101A/K102A, Delta YRKL, 4A, PTAP, 4A+PTAP, and YPDL) when expressed alone from cloned cDNAs were only cytoplasmic, whereas the wild-type M1 expressed alone was both nuclear and cytoplasmic as expected. These results show that the nuclear translocation function provided by the positively charged residues within the NLS motif does not play a critical role in influenza virus replication. Furthermore, these sequences of H6 domain can be replaced by late (L) domain motifs and therefore may provide a function similar to that of the L domains of other negative-strand RNA and retroviruses.

MeSH Terms
Amino Acid Motifs Amino Acid Sequence Animals Cell Line Cell Nucleus/metabolism Cytoplasm/metabolism Dogs Humans Influenza A virus/metabolism,physiology Microscopy, Electron Molecular Sequence Data Mutation Nuclear Localization Signals/chemistry,genetics,metabolism Structure-Activity Relationship Viral Matrix Proteins/chemistry,genetics,metabolism Virus Assembly Virus Replication
Chemicals
Nuclear Localization Signals Viral Matrix Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Hui Eric Ka-Wai
Department of Microbiology, Immunology and Molecular Genetics, Jonsson Comprehensive Cancer Center, Molecular Biology Institute, UCLA School of Medicine, Los Angeles, California 90095-1747, USA.
Barman Subrata
Yang Tae Yong
Nayak Debi P
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/jvi.77.12.7078-7092.2003
Published
2003-06-00
Pages
7078-92
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC156155
Subset
IM
Grants
NIAID NIH HHS · R01 AI016348 · United States
NIAID NIH HHS · R01 AI041681 · United States
NIAID NIH HHS · AI 16348 · United States
NIAID NIH HHS · AI 41681 · United States
Corrections
RetractionIn
PMID: 17005709
External Links
PubMed source DOI: 10.1128/jvi.77.12.7078-7092.2003 PMC Full Text View journal details
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