scholarly journals Influenza B Virus BM2 Protein Is Transported through the trans-Golgi Network as an Integral Membrane Protein

2003 ◽  
Vol 77 (19) ◽  
pp. 10630-10637 ◽  
Author(s):  
Shinji Watanabe ◽  
Masaki Imai ◽  
Yoshiro Ohara ◽  
Takato Odagiri
Keyword(s):  
Plasma Membrane ◽  
Membrane Protein ◽  
Matrix Protein ◽  
Integral Membrane Protein ◽  
Influenza B Virus ◽  
Influenza B ◽  
Hydrophobic Domain ◽  
Cytoplasmic Transport ◽  
B Virus ◽  
Golgi Network

ABSTRACT A bicistronic mRNA transcribed from the influenza B virus RNA segment 7 encodes two viral proteins, matrix protein M1 and uncharacterized small protein BM2. In the present study, we focused on the cytoplasmic transport and cellular membrane association of BM2. Immunofluorescence studies of virus-infected cells indicated that BM2 accumulated at the Golgi apparatus immediately after synthesis and then was transported to the plasma membrane through the trans-Golgi network. Localization of a set of BM2 deletion mutants revealed that the N-terminal half of BM2 (residues 2 to 50) was crucial for its transport; in particular, the deletion of residues 2 to 23, deduced to be a transmembrane domain, resulted in diffused distribution of the protein throughout the entire cell. Sucrose gradient flotation and biochemical analyses of the membrane showed that BM2 was tightly associated with cellular membranes as an integral membrane protein. Oligomerization of BM2 was demonstrated by coprecipitation of differentially epitope-tagged BM2 proteins. Taken together, these results strongly suggest that BM2 is integrated into the plasma membrane at the N-terminal hydrophobic domain as fourth membrane protein, in addition to hemagglutinin, neuraminidase, and NB, of the influenza B virus.

scholarly journals Determination of the orientation of an integral membrane protein and sites of glycosylation by oligonucleotide-directed mutagenesis: influenza B virus NB glycoprotein lacks a cleavable signal sequence and has an extracellular NH2-terminal region.

1986 ◽  
Vol 6 (12) ◽  
pp. 4317-4328 ◽  
Author(s):  
M A Williams ◽  
R A Lamb
Keyword(s):  
Membrane Protein ◽  
Cell Surface ◽  
Signal Sequence ◽  
Integral Membrane Protein ◽  
Terminal Region ◽  
Influenza B Virus ◽  
Influenza B ◽  
Directed Mutagenesis ◽  
Cell Fractionation ◽  
B Virus

The membrane orientation of the NB protein of influenza B virus, a small (Mr, approximately 18,000) glycoprotein with a single internal hydrophobic domain, was investigated by biochemical and genetic means. Cell fractionation and protein solubility studies indicate NB is an integral membrane protein, and NB has been shown to be a dimer under nonreducing conditions. Treatment of infected-cell surfaces with proteinase K and endoglycosidase F and immunoprecipitation with a site-specific antibody suggests that the 18-amino-acid NH2-terminal region of NB is exposed at the cell surface. Oligonucleotide-directed mutagenesis to eliminate each of the four potential sites of N-linked glycosylation and expression of the mutant NB proteins in eucaryotic cells suggest that the two sites adjacent to the NH2 terminus are glycosylated. This provides further evidence that NB, which lacks a cleavable NH2-terminal signal sequence, has an exposed NH2 terminus at the cell surface.

scholarly journals Influenza B virus BM2 protein is an oligomeric integral membrane protein expressed at the cell surface

Virology ◽  
2003 ◽  
Vol 306 (1) ◽  
pp. 7-17 ◽  
Author(s):  
Reay G Paterson ◽  
Makoto Takeda ◽  
Yuki Ohigashi ◽  
Lawrence H Pinto ◽  
Robert A Lamb
Keyword(s):  
Membrane Protein ◽  
Cell Surface ◽  
Integral Membrane Protein ◽  
Influenza B Virus ◽  
Influenza B ◽  
B Virus

Determination of the orientation of an integral membrane protein and sites of glycosylation by oligonucleotide-directed mutagenesis: influenza B virus NB glycoprotein lacks a cleavable signal sequence and has an extracellular NH2-terminal region

1986 ◽  
Vol 6 (12) ◽  
pp. 4317-4328
Author(s):  
M A Williams ◽  
R A Lamb
Keyword(s):  
Membrane Protein ◽  
Cell Surface ◽  
Signal Sequence ◽  
Integral Membrane Protein ◽  
Terminal Region ◽  
Influenza B Virus ◽  
Influenza B ◽  
Directed Mutagenesis ◽  
Cell Fractionation ◽  
B Virus

The membrane orientation of the NB protein of influenza B virus, a small (Mr, approximately 18,000) glycoprotein with a single internal hydrophobic domain, was investigated by biochemical and genetic means. Cell fractionation and protein solubility studies indicate NB is an integral membrane protein, and NB has been shown to be a dimer under nonreducing conditions. Treatment of infected-cell surfaces with proteinase K and endoglycosidase F and immunoprecipitation with a site-specific antibody suggests that the 18-amino-acid NH2-terminal region of NB is exposed at the cell surface. Oligonucleotide-directed mutagenesis to eliminate each of the four potential sites of N-linked glycosylation and expression of the mutant NB proteins in eucaryotic cells suggest that the two sites adjacent to the NH2 terminus are glycosylated. This provides further evidence that NB, which lacks a cleavable NH2-terminal signal sequence, has an exposed NH2 terminus at the cell surface.

scholarly journals VIP21, a 21-kD membrane protein is an integral component of trans-Golgi-network-derived transport vesicles.

1992 ◽  
Vol 118 (5) ◽  
pp. 1003-1014 ◽  
Author(s):  
T V Kurzchalia ◽  
P Dupree ◽  
R G Parton ◽  
R Kellner ◽  
H Virta ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Membrane Protein ◽  
Integral Membrane Protein ◽  
Cellular Membrane ◽  
Integral Membrane Proteins ◽  
Membrane Domains ◽  
Transport Vesicles ◽  
Vesicular Carriers ◽  
Molecular Machinery ◽  
Golgi Network

In simple epithelial cells, apical and basolateral proteins are sorted into separate vesicular carriers before delivery to the appropriate plasma membrane domains. To dissect the putative sorting machinery, we have solubilized Golgi-derived transport vesicles with the detergent CHAPS and shown that an apical marker, influenza haemagglutinin (HA), formed a large complex together with several integral membrane proteins. Remarkably, a similar set of CHAPS-insoluble proteins was found after solubilization of a total cellular membrane fraction. This allowed the cloning of a cDNA encoding one protein of this complex, VIP21 (Vesicular Integral-membrane Protein of 21 kD). The transiently expressed protein appeared on the Golgi-apparatus, the plasma membrane and vesicular structures. We propose that VIP21 is a component of the molecular machinery of vesicular transport.

scholarly journals Characterization of the Nucleocytoplasmic Shuttle of the Matrix Protein of Influenza B Virus

2014 ◽  
Vol 88 (13) ◽  
pp. 7464-7473 ◽  
Author(s):  
S. Cao ◽  
J. Jiang ◽  
J. Li ◽  
Y. Li ◽  
L. Yang ◽  
...  
Keyword(s):  
Matrix Protein ◽  
Influenza B Virus ◽  
Influenza B ◽  
Nucleocytoplasmic Shuttle ◽  
B Virus ◽  
The Matrix

scholarly journals Analysis of the desialidation process of the haemagglutinin protein of influenza B virus: the host-dependent desialidation step

2002 ◽  
Vol 83 (7) ◽  
pp. 1729-1734 ◽  
Author(s):  
C. Luo ◽  
E. Nobusawa ◽  
K. Nakajima
Keyword(s):  
Cell Surface ◽  
Mdck Cells ◽  
Influenza B Virus ◽  
Influenza B ◽  
Dependent Manner ◽  
Cos Cells ◽  
Trans Golgi Network ◽  
B Virus ◽  
Golgi Network ◽  
Ha Protein

It was reported previously that haemadsorption by the haemagglutinin (HA) protein of influenza B virus required that the protein must undergo desialidation. When MDCK and COS cells were infected with influenza B/Kanagawa/73 virus in the presence of a neuraminidase (NA) inhibitor, Zanamivir, haemadsorption on MDCK cells was inhibited but that on COS cells was not. The activity of the NA protein of the two types of infected cells was similar and both were inhibited by Zanamivir in a dose-dependent manner. A comparison of the desialidation of the HA protein was made on MDCK and COS cells in the presence of bacterial NA and both cells were found to have similar sensitivity. On the accumulation of the HA and NA proteins in the trans-Golgi network of MDCK cells by means of low-temperature treatment, desialidation of the HA protein in the presence of Zanamivir was detected by two-dimensional gel electrophoresis. Because this agent was reported to be unable to penetrate cells, these data suggest that, in MDCK cells, desialidation of the HA protein occurs on the cell surface but, in COS cells, the HA and NA proteins might accumulate in the trans-Golgi network, thus allowing NA desialidation before their migration to the cell surface.

scholarly journals Human anti-p68 autoantibodies recognize a common epitope of U1 RNA containing small nuclear ribonucleoprotein and influenza B virus.

1990 ◽  
Vol 171 (3) ◽  
pp. 819-829 ◽  
Author(s):  
H H Guldner ◽  
H J Netter ◽  
C Szostecki ◽  
E Jaeger ◽  
H Will
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Matrix Protein ◽  
Autoimmune Response ◽  
Influenza B Virus ◽  
Influenza B ◽  
Sequence Motif ◽  
Small Nuclear Ribonucleoprotein ◽  
B Virus ◽  
Nuclear Ribonucleoprotein

Autoantibodies from patients with systemic rheumatic diseases were used to map antigenic sites on the 68-kD autoantigen (p68) associated with (U1)RNA-containing small nuclear ribonucleoprotein (snRNP) particles. With truncated recombinant fusion proteins and synthetic peptides, a subset of anti-p68 autoantibodies was found to recognize the amino acid sequence motif Glu-Arg-Lys-Arg-Arg (ERKRR). To investigate the possible involvement of epitopes shared by microbial antigens and host self-components in initiation of autoimmunity (molecular mimicry), a sequence data bank was screened for proteins containing an amino acid motif identical or related to ERKRR. The identical motif was found on the M1 matrix protein of influenza B viruses, and affinity-purified human anti-ERKRR autoantibodies recognized this epitope also in the viral amino acid sequence context. The common epitope recognized by human autoantibodies suggests that influenza B virus infection may play a role in initiation of the anti-p68 and anti-(U1)RNP autoimmune response.

scholarly journals Palmitoylation is required for intracellular trafficking of influenza B virus NB protein and efficient influenza B virus growth in vitro

2014 ◽  
Vol 95 (6) ◽  
pp. 1211-1220 ◽  
Author(s):  
Andrew Demers ◽  
Zhiguang Ran ◽  
Qiji Deng ◽  
Dan Wang ◽  
Brody Edman ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Lipid Rafts ◽  
Transmembrane Domain ◽  
Influenza Viruses ◽  
Influenza B Virus ◽  
Apical Plasma Membrane ◽  
Influenza B ◽  
Virus Growth ◽  
B Virus

All influenza viruses bud and egress from lipid rafts within the apical plasma membrane of infected epithelial cells. As a result, all components of progeny virions must be transported to these lipid rafts for assembly and budding. Although the mechanism of transport for other influenza proteins has been elucidated, influenza B virus (IBV) glycoprotein NB subcellular localization and transport are not understood completely. To address the aforementioned properties of NB, a series of trafficking experiments were conducted. Here, we showed that NB co-localized with markers specific for the endoplasmic reticulum (ER) and Golgi region. The data from chemical treatment of NB-expressing cells by Brefeldin A, a fungal antibiotic and a known chemical inhibitor of the protein secretory pathway, further confirmed that NB is transported through the ER–Golgi pathway as it restricted NB localization to the perinuclear region. Using NB deletion mutants, the hydrophobic transmembrane domain was identified as being required for NB transport to the plasma membrane. Furthermore, palmitoylation was also required for transport of NB to the plasma membrane. Systematic mutation of cysteines to serines in NB demonstrated that cysteine 49, likely in a palmitoylated form, is also required for transport to the plasma membrane. Surprisingly, further analysis demonstrated that in vitro replication of NBC49S mutant virus was delayed relative to the parental IBV. The results demonstrated that NB is the third influenza virus protein to have been shown to be palmitoylated and together these findings may aid in future studies aimed at elucidating the function of NB.

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