In Vitro Assembly of the Murine Coronavirus Membrane Protein E1

Rottier, Peter J. M.; Brandenburg, Dorothee; Armstrong, J. Scott; Zeijst, Ben van der; Warren, Graham

doi:10.1007/978-1-4615-9373-7_5

Cited by 7 publications

(12 citation statements)

References 35 publications

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“…The exterior surface of the microsomes is derived from the cytoplasmic side of the ER (27,32). Protease protection assays have been used to determine the topology of a number of membrane proteins (33)(34)(35)(36)(37). Domains protected from protease digestion by microsomes are located in the interior of the microsome or within the microsomal membrane itself and thus are ectodomains or transmembrane domains, respectively.…”

Section: Resultsmentioning

confidence: 99%

Determination of the Transmembrane Topology of Herpes Simplex Virus Type 1 Glycoprotein K (gK)

Holland

1997

Journal of Biological Chemistry

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Herpes simplex virus type 1 glycoprotein K (gK) plays an essential role in viral replication and cell fusion. gK is a very hydrophobic membrane protein that contains a signal sequence and several hydrophobic regions. It has been shown that mutations inducing cell fusion map to two distinct domains of gK, suggesting that these domains are functionally important. To understand the transmembrane topology of gK and the localization of these functional domains, we constructed a set of gK deletion, insertion, and truncation mutants and expressed these by in vitro translation in the presence of microsomal membranes. The transmembrane topology of gK was determined by examination of the post-translational processing and protease sensitivity of the mutant proteins. Our data demonstrate that gK contains three transmembrane domains (amino acids 125-139, 226-239, and 311-325). Another hydrophobic domain (amino acids 241-265), which is relatively less hydrophobic and much longer compared with the transmembrane sequences, is located in the extracellular loop. The analysis showed that the domains containing syncytial mutations are both ectodomains. They may interact with each other to form a complex tertiary structure that is critical for the biological function of gK.

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Section: Resultsmentioning

confidence: 99%

Determination of the Transmembrane Topology of Herpes Simplex Virus Type 1 Glycoprotein K (gK)

Holland

1997

Journal of Biological Chemistry

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“…Previous studies have demonstrated that saponin treatment will make not only the plasma membrane but also intracellular membranes permeable to proteins (Ohtsuki et al, 1978;Castle & Palade, 1978;Rottier et al, 1984;Morgan & Peters, 1985). There is, however, little quantitative data in the literature on the interaction of any specific saponin with different intracellular membranes in one and the same cell type.…”

Section: Discussionmentioning

confidence: 99%

“…Treatment with bacterial toxins (Ahnert-Hilger et al, 1985;McEwen & Arion, 1985), the polyene antibiotic filipin (Jorgensen & Nordlie, 1980;Gankema et al, 1981), or saponins (at low concentrations) (Fiskum et al, 1980;Hirata & Koga, 1982;Burgess et al, 1983) has been widely used to permeabilize the plasma membrane of different cells. Saponins have also been used (at high concentrations) in immunocytochemistry to permeabilize intracellular membranes (Ohtsuki et al, 1978;Willingham et al, 1978;Saraste & Hedman, 1983) and to permeabilize membrane vesicles derived from intracellular membranes (Castle & Palade, 1978;Rottier et al, 1984).…”

Section: Introductionmentioning

confidence: 99%

Differential permeabilization of membranes by saponin treatment of isolated rat hepatocytes. Release of secretory proteins

Wassler

Jonasson

Persson

et al. 1987

Biochemical Journal

104

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Monolayer cultures of rat hepatocytes were treated with increasing concentrations of saponin (prepared from Gypsophila plants) for 30 min at 6 degrees C. Differential permeabilization of the intracellular membranes could be demonstrated: at 0.040 mg of saponin/ml the plasma membrane was permeabilized, as assessed by the release of 50% of the total cellular amount of lactate dehydrogenase, and at 0.20 mg/ml the endoplasmic reticulum was permeabilized, as measured by the release of 50% of pulse-35S-labelled albumin. The Golgi complex was permeabilized at an intermediate saponin concentration, as indicated by the release of homogeneously 35S-labelled albumin; about half the intracellular albumin is located in this organelle. At 1.0 up to 5.0 mg of saponin/ml 90-95% of the radioactively labelled albumin was released. Even at 5.0 mg/ml less than 10% of the membrane of the endoplasmic reticulum was solubilized, as judged by the degree of release of a membrane-bound enzyme specific for this organelle. These results demonstrate the usefulness of saponin as a tool for investigating the interior of different intracellular compartments.

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“…Through the interaction with M protein, S protein is incorporated into the viral envelope [17,18] and the mature virions are released from the cells. These studies suggest that coronavirus M protein [19,20] plays a crucial role in the assembly of virus particles. Like other coronaviruses, SARS-CoV assembles at and buds into the lumen of the endoplasmic reticulum-Golgi intermediate compartment [21].…”

Section: Introductionmentioning

confidence: 89%

Interactions between M protein and other structural proteins of severe, acute respiratory syndrome-associated coronavirus

Hsieh

Chen

et al. 2008

J Biomed Sci

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Severe acute respiratory syndrome-associated coronavirus (SARS-CoV) structural proteins (S, E, M, and NC) localize in different subcellular positions when expressed individually. However, SARS-CoV M protein is co-localized almost entirely with S, E, or NC protein when co-expressed in the cells. On the other hand, only partial co-localization was observed when S and E, S and NC, or E and NC were co-expressed in the cells. Interactions between SARS-CoV M and other structural proteins but not interactions between S and E, S and NC, or E and NC were further demonstrated by co-immunoprecipitation assay. These results indicate that SARS-CoV M protein, similar to the M proteins of other coronaviruses, plays a pivotal role in virus assembly. The cytoplasmic C-terminus domain of SARS-CoV M protein was responsible for binding to NC protein. Multiple regions of M protein interacted with E and S proteins. A model for the interactions between SARS-CoV M protein and other structural proteins is proposed. This study helps us better understand protein-protein interactions during viral assembly of SARS-CoV.

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In Vitro Assembly of the Murine Coronavirus Membrane Protein E1

Cited by 7 publications

References 35 publications

Determination of the Transmembrane Topology of Herpes Simplex Virus Type 1 Glycoprotein K (gK)

Determination of the Transmembrane Topology of Herpes Simplex Virus Type 1 Glycoprotein K (gK)

Differential permeabilization of membranes by saponin treatment of isolated rat hepatocytes. Release of secretory proteins

Interactions between M protein and other structural proteins of severe, acute respiratory syndrome-associated coronavirus

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