Preferential Initiation at the Second AUG of the Measles Virus F mRNA: A Role for the Long Untranslated Region

Cathomen, Toni; Buchholz, Christian J.; Spielhofer, Pius; Cattaneo, Roberto

doi:10.1006/viro.1995.0075

Cited by 87 publications

(71 citation statements)

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“…Plasmids, Mutagenesis, and Receptor-specific Cell-to-Cell Fusion Assay-All H-protein mutants were generated in the pCG-H plasmid (19) by the QuikChange site-directed mutagenesis (Stratagene). The integrity of the clone was confirmed by sequencing the H-protein gene in the vicinity of the mutation.…”

Section: Methodsmentioning

confidence: 99%

Dynamic Interaction of the Measles Virus Hemagglutinin with Its Receptor Signaling Lymphocytic Activation Molecule (SLAM, CD150)

Navaratnarajah

Vongpunsawad

Oezguen

et al. 2008

Journal of Biological Chemistry

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The interaction of measles virus with its receptor signaling lymphocytic activation molecule (SLAM) controls cell entry and governs tropism. We predicted potential interface areas of the measles virus attachment protein hemagglutinin to begin the investigation. We then assessed the relevance of individual amino acids located in these areas for SLAM-binding and SLAM-dependent membrane fusion, as measured by surface plasmon resonance and receptor-specific fusion assays, respectively. These studies identified one hemagglutinin protein residue, isoleucine 194, which is essential for primary binding. The crystal structure of the hemagglutinin-protein localizes Ile-194 at the interface of propeller blades 5 and 6, and our data indicate that a small aliphatic side chain of residue 194 stabilizes a protein conformation conducive to binding. In contrast, a quartet of residues previously shown to sustain SLAM-dependent fusion is not involved in binding. Instead, our data prove that after binding, this quartet of residues on propeller blade 5 conducts conformational changes that are receptor-specific. Our study sets a structure-based stage for understanding how the SLAM-elicited conformational changes travel through the H-protein ectodomain before triggering fusion protein unfolding and membrane fusion.Measles is a leading cause of childhood morbidity and mortality in developing countries (1). The immune suppression that accompanies infection with wild-type measles viruses (MV) 2 exposes individuals to secondary infections causing most of the fatalities associated with the disease (2). Live attenuated MV vaccines have effectively reduced the morbidity and mortality of measles world-wide (3). The difference in pathology between the wild-type and vaccine strains is in part explained by the receptors used for cell entry. Wild-type MV uses the signaling lymphocyte activation molecule (SLAM, CD150) (4 -7), while the vaccine strain has gained the ability to also use the ubiquitous protein CD46 (8 -10). SLAM-targeting accounts for lymphotropism and is central to understanding MV tropism and disease progression.Receptor attachment of MV and the other morbilliviruses is mediated by hemagglutinin (H) homodimers. Unlike other members of the Paramyxoviridae, the morbillivirus H-proteins do not have neuraminidase activity. Upon receptor attachment, H induces conformational changes in the trimeric fusion (F) protein, resulting in the fusion of viral and cellular membranes (11,12). MV H is a 617-amino acid type II transmembrane glycoprotein, which is comprised of an N-terminal cytoplasmic tail, a membrane-spanning domain, and an extracellular stalk region connected to a C-terminal globular head (13).Previously, we generated a three-dimensional model of the extracellular domain of the MV H-protein based on the crystal structure of Newcastle Disease Virus (NDV) HN (hemagglutinin neuraminidase) protein (sequence identity of 12%), a related paramyxovirus (14,15). This model predicts that the MV H ectodomain has a 6-blade propeller structure....

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Section: Methodsmentioning

confidence: 99%

Dynamic Interaction of the Measles Virus Hemagglutinin with Its Receptor Signaling Lymphocytic Activation Molecule (SLAM, CD150)

Navaratnarajah

Vongpunsawad

Oezguen

et al. 2008

Journal of Biological Chemistry

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“…Purified PCR products were mixed in a molar ratio of 1 : 1 : 1 and hybridization was carried out as reported previously (Köhl et al, 2004). After amplifying the hybrid genes, the PCR products were digested with BamHI and XbaI and the fragments ligated into the vector pCG1 (Cathomen et al, 1995). For site-directed mutagenesis for generation of E1-C/A, E1-C/S, Flag-E1-K, Flag-E1-R and Flag-E1-KR, modified sense and antisense primers were used.…”

Section: Methodsmentioning

confidence: 99%

Formation of bovine viral diarrhea virus E1–E2 heterodimers is essential for virus entry and depends on charged residues in the transmembrane domains

Ronecker

Zimmer

Herrler

et al. 2008

Journal of General Virology

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The envelope of bovine viral diarrhea virus (BVDV) contains the glycoproteins E rns , E1 and E2. Complementation of a recombinant vesicular stomatitis virus (VSV) with BVDV glycoproteins resulted in infectious pseudotyped viruses. To elucidate the specific role of each of the single envelope glycoproteins during viral entry, pseudotypes were generated bearing the BVDV envelope proteins in different combinations. Pseudoviruses that contained E1 and E2 but not E rns were infectious, indicating that E rns is dispensable for virus entry. VSV/BVDV pseudotypes with chimeric proteins (the ectodomain of the BVDV glycoprotein and the transmembrane domain of the VSV-G protein) were not infectious. The fact that E1-E2 heterodimers were not detected if one of the proteins was chimeric indicated that the heterodimers are crucial for BVDV entry. It was shown by site-directed mutagenesis that the charged amino acids in the transmembrane domains of BVDV E1 (lysine and arginine) and the charged amino acid in the transmembrane domain of E2 (arginine) play a key role in heterodimer formation. Pseudoviruses bearing the mutation E2-R/A, where the charged amino acid was substituted by alanine, were not infectious, supporting the hypothesis that E1-E2 heterodimers are essential for BVDV entry.

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“…Plasmid Construction-Parental plasmids for mutagenesis and all experiments were pCG-H Edm and pCG-F Edm encoding MV-Edm H and F under the control of the cytomegalovirus promoter (28). Site-directed mutagenesis was performed using the quick change system (Stratagene) and mutant constructs confirmed by DNA sequencing and Western analysis.…”

Section: Methodsmentioning

confidence: 99%

Measles Virus Envelope Glycoproteins Hetero-oligomerize in the Endoplasmic Reticulum

Plemper

Hammond

Cattaneo

2001

Journal of Biological Chemistry

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123

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The endoplasmic reticulum (ER) was investigated as the initial oligomerization site for the envelope glycoproteins H and F of measles virus (MV), a clinically relevant member of the Paramyxoviridae family, and consequences of this interaction for viral replication were studied. Both proteins were tagged at their cytosolic tails with RRR and KKXX motifs, respectively, resulting in their efficient retention in the ER. Cotransfection of the retained constructs with transport competent MV glycoproteins revealed a dominant negative effect on their biological activity indicating intracellular complex formation and thus retention. Pulse-chase analysis and co-immunoprecipitation experiments demonstrated that this effect is based on both homo-and hetero-oligomerization in the ER. Recombinant viruses additionally expressing ERretained F showed an altered cytopathic phenotype accompanied by greatly reduced particle release. Similar mutant viruses additionally expressing ERretained H could not be rescued indicating an even greater negative effect of this protein on virus viability. Our study suggests that both homo-and heterooligomerization of MV glycoproteins occur in the ER and that these events are of significance for early steps of particle assembly.

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Preferential Initiation at the Second AUG of the Measles Virus F mRNA: A Role for the Long Untranslated Region

Cited by 87 publications

References 0 publications

Dynamic Interaction of the Measles Virus Hemagglutinin with Its Receptor Signaling Lymphocytic Activation Molecule (SLAM, CD150)

Dynamic Interaction of the Measles Virus Hemagglutinin with Its Receptor Signaling Lymphocytic Activation Molecule (SLAM, CD150)

Formation of bovine viral diarrhea virus E1–E2 heterodimers is essential for virus entry and depends on charged residues in the transmembrane domains

Measles Virus Envelope Glycoproteins Hetero-oligomerize in the Endoplasmic Reticulum

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