The
            <i>in situ</i>
            spatial arrangement of the influenza A virus matrix protein M1 assessed by tritium bombardment

Shishkov, A.V.; Goldanskii, V. I.; Baratova, Ludmila A.; Федорова, Н. В.; Ksenofontov, Alexander L.; Жирнов, О. П.; Galkin, Andrey

doi:10.1073/pnas.96.14.7827

Cited by 61 publications

(51 citation statements)

References 19 publications

(16 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although the structure of the C-terminal sequence of the proposed zinc finger motif in L9 of influenza virus M1 has not been defined, two cysteines in the middle of the H9 domain are in the a-helix conformation (Fig. 1) (Sha & Luo, 1997;Shishkov et al, 1999;Arzt et al, 2001;Harris et al, 2001;Okada et al, 2003). Therefore, these residues are unlikely to form a coordinate complex with zinc.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Conserved cysteine and histidine residues in the putative zinc finger motif of the influenza A virus M1 protein are not critical for influenza virus replication

Hui¹,

Ralston²,

Judd³

et al. 2003

Journal of General Virology

View full text Add to dashboard Cite

The influenza virus matrix protein (M1) possesses a cysteine and histidine (CCHH) motif in the helix 9 (H9) and adjacent region ( 148 CATCEQIADSQHRSH 162 ). The CCHH motif has been proposed as a putative zinc finger motif and zinc-binding activity has been implicated in virus uncoating as well as transcription inhibition and mRNA regulation. The function of the CCHH motif in the influenza virus life cycle was investigated by site-directed mutagenesis (alanine replacement) and by rescuing mutant viruses by reverse genetics. Mutant viruses containing an alanine replacement of the cysteine and histidine residues, either individually or in combination, were seen to exhibit wt phenotype in multiple virus growth cycles and plaque morphology. In addition, synthetic peptides containing the putative zinc finger motif did not inhibit virus replication in MDCK cells. However, mutation of Ala 155 in H9 was lethal for rescuing infectious virus.These data show that the CCHH motif does not provide a critical function in the influenza virus life cycle in cell culture and that the zinc-binding function may not be involved in virus biology. However, the lethal phenotype of the Ala 155 mutation shows that the H9 region of M1 provides some other critical function(s) in virus replication.

show abstract

Section: Discussionmentioning

confidence: 99%

“…The structure of the N terminus (aa 2-164) of M1 has been determined by crystallization, X-ray diffraction, circular dichroism and tritium bombardment (Sha & Luo, 1997;Shishkov et al, 1999;Arzt et al, 2001;Harris et al, 2001). It contains two spatially distinct domains, N and M (Fig.…”

Section: Introductionmentioning

confidence: 99%

Conserved cysteine and histidine residues in the putative zinc finger motif of the influenza A virus M1 protein are not critical for influenza virus replication

Hui¹,

Ralston²,

Judd³

et al. 2003

Journal of General Virology

View full text Add to dashboard Cite

show abstract

“…This approach has been used successfully to probe directly the accessible surfaces of proteins in macromolecular assemblies such as those of the plant viruses tobacco mosaic virus (TMV) (genus Tobamovirus) (16) and potato virus X (PVX) (genus Potexvirus) (7), the ribosome (1), and membrane structures (30,34).…”

mentioning

confidence: 99%

In Situ Spatial Organization of Potato Virus A Coat Protein Subunits as Assessed by Tritium Bombardment

Baratova

Ефимов

Добров

et al. 2001

J Virol

Self Cite

View full text Add to dashboard Cite

Potato virus A (PVA) particles were bombarded with thermally activated tritium atoms, and the intramolecular distribution of the label in the amino acids of the coat protein was determined to assess their in situ steric accessibility. This method revealed that the N-terminal 15 amino acids of the PVA coat protein and a region comprising amino acids 27 to 50 are the most accessible at the particle surface to labeling with tritium atoms. A model of the spatial arrangement of the PVA coat protein polypeptide chain within the virus particle was derived from the experimental data obtained by tritium bombardment combined with predictions of secondary-structure elements and the principles of packing ␣-helices and ␤-structures in proteins. The model predicts three regions of tertiary structure: (i) the surface-exposed N-terminal region, comprising an unstructured N terminus of 8 amino acids and two ␤-strands, (ii) a C-terminal region including two ␣-helices, as well as three ␤-strands that form a two-layer structure called an abCd unit, and (iii) a central region comprising a bundle of four ␣-helices in a fold similar to that found in tobacco mosaic virus coat protein. This is the first model of the three-dimensional structure of a potyvirus coat protein.

show abstract

“…1 F and G). Although there is no X-ray structure currently available for the C domain of FLUA-M1, it is predicted to contain four α-helices (39,40), exactly what is observed in the ISAV-M1 structure. The high α-helical content of the FLUA-M1 C domain was also confirmed by a number of solution studies (15,39).…”

Section: Resultsmentioning

confidence: 99%

“…Although there is no X-ray structure currently available for the C domain of FLUA-M1, it is predicted to contain four α-helices (39,40), exactly what is observed in the ISAV-M1 structure. The high α-helical content of the FLUA-M1 C domain was also confirmed by a number of solution studies (15,39). Additional evidence suggesting that ISAV-M1 and FLUA-M1 share similar structure and function includes: (i) The elbowshape of ISAV-M1 closely resembles that of virion-associated FLUA-M1 molecules seen in negative-staining EM images (18); (ii) both ISAV-M1 and FLUA-M1 exist as monomers in solution (15); and (iii) ISAV-M1 demonstrates an inherent membrane association activity similar to that of the FLUA-M1 (18).…”

Section: Resultsmentioning

confidence: 99%

The crystal structure of an orthomyxovirus matrix protein reveals mechanisms for self-polymerization and membrane association

Tao¹

2017

J Immunol Tech Infect Dis

View full text Add to dashboard Cite

Many enveloped viruses encode a matrix protein. In the influenza A virus, the matrix protein M1 polymerizes into a rigid protein layer underneath the viral envelope to help enforce the shape and structural integrity of intact viruses. The influenza virus M1 is also known to mediate virus budding as well as the nuclear export of the viral nucleocapsids and their subsequent packaging into nascent viral particles. Despite extensive studies on the influenza A virus M1 (FLUA-M1), only crystal structures of its N-terminal domain are available. Here we report the crystal structure of the full-length M1 from another orthomyxovirus that infects fish, the infectious salmon anemia virus (ISAV). The structure of ISAV-M1 assumes the shape of an elbow, with its N domain closely resembling that of the FLUA-M1. The C domain, which is connected to the N domain through a flexible linker, is made of four α-helices packed as a tight bundle. In the crystal, ISAV-M1 monomers form infinite 2D arrays with a network of interactions involving both the N and C domains. Results from liposome flotation assays indicated that ISAV-M1 binds membrane via electrostatic interactions that are primarily mediated by a positively charged surface loop from the N domain. Cryoelectron tomography reconstruction of intact ISA virions identified a matrix protein layer adjacent to the inner leaflet of the viral membrane. The physical dimensions of the virion-associated matrix layer are consistent with the 2D ISAV-M1 crystal lattice, suggesting that the crystal lattice is a valid model for studying M1-M1, M1-membrane, and M1-RNP interactions in the virion.ll members of the Orthomyxoviridae family, including the influenza viruses A-D, thogotovirus, and isavirus, encode a matrix protein called M1. In the influenza A virus, M1 is produced by a colinear transcript made from the gene segment 7 (1). As one of the most abundantly made viral proteins, the influenza A virus M1 plays multiple roles during the virus life cycle. Upon viral entry, the acidification of the viral interior in the endosome weakens the interaction between M1 and the viral ribonucleoprotein complexes (vRNPs), thus allowing M1-free vRNPs to be imported to the nucleus for viral RNA replication and transcription (2, 3). As infection proceeds, newly synthesized M1 enters the nucleus to mediate the nuclear export of nascent vRNPs. A "daisy-chain" complex is formed with the vRNP binding to the C-terminal domain of M1 (4) and the N-terminal domain of M1 interacting with the nuclear export protein (NEP), also called NS2. Through its nuclear export signal (NES), NEP is specifically recognized by the cellular exportin Crm1, which then facilitates the transport of vRNPs across the nuclear membrane to the cytoplasm in a RanGTP-dependent manner (5). It has been shown that M1 binding to vRNP in the nucleus is able to block mRNA transcription (4, 6). In the cytosol, M1 interacts with the cytoplasmic tails of the glycoproteins HA and NA. Such interactions promote M1 association with lipid raft membranes and t...

show abstract

The in situ spatial arrangement of the influenza A virus matrix protein M1 assessed by tritium bombardment

Cited by 61 publications

References 19 publications

Conserved cysteine and histidine residues in the putative zinc finger motif of the influenza A virus M1 protein are not critical for influenza virus replication

Conserved cysteine and histidine residues in the putative zinc finger motif of the influenza A virus M1 protein are not critical for influenza virus replication

In Situ Spatial Organization of Potato Virus A Coat Protein Subunits as Assessed by Tritium Bombardment

The crystal structure of an orthomyxovirus matrix protein reveals mechanisms for self-polymerization and membrane association

Contact Info

Product

Resources

About