In vitro dissection of the membrane and RNP binding activities of influenza virus M1 protein.
Spontaneous proteolysis of influenza virus M1 protein during crystallisation has defined an N-terminal domain of amino acids 1--164. Full-length M1, the N-terminal domain, and the C-terminal part of M1 (residues 165--252) were produced in Escherichia coli. In vitro tests showed that only full-length M1 and its N-terminal domain bind to negatively charged liposomes and that only full-length M1 and its C-terminal part bind to RNP. However, only full-length M1 had transcription inhibition activity. Several independent experimental approaches indicate that in vitro transcription inhibition occurs through polymerisation/aggregation of M1 onto RNP, or of M1 onto M1 already bound to RNP, rather than by binding to a specific active site on the nucleoprotein or the polymerase. The structure/function of influenza virus M1 will be compared with that of the Ebola virus matrix protein, VP40.
Baudin
F
F
EMBL Grenoble Outstation, B.P. 156, 38042 Grenoble Cedex 9, France.
Petit
I
I
Weissenhorn
W
W
Ruigrok
R W
RW
D029043:Ebolavirus / Q000737:chemistry; D008081:Liposomes / Q000378:metabolism*; D008854:Microscopy, Electron; D009154:Mutation / Q000235:genetics; D019913:Nuclear Localization Signals / Q000235:genetics; D009975:Orthomyxoviridae / Q000737:chemistry; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012261:Ribonucleoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D012965:Sodium Chloride / Q000494:pharmacology; D012995:Solubility / Q000187:drug effects; D055672:Static Electricity; D014158:Transcription, Genetic; D014763:Viral Matrix Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure
D008081:Liposomes; C031540:M-protein, influenza virus; C488936:M1 protein, Influenza A virus; D019913:Nuclear Localization Signals; D010446:Peptide Fragments; D012261:Ribonucleoproteins; D014763:Viral Matrix Proteins; D012965:Sodium Chloride