Trudy Morrison - Academia.edu (original) (raw)

Papers by Trudy Morrison

Journal of Virology, Dec 1, 1975

We have studied the relationships among the polypeptides of Newcastle disease virus by using both... more We have studied the relationships among the polypeptides of Newcastle disease virus by using both kinetic and tryptic peptide analyses. The results of our tryptic peptide analyses suggest that there are at least six unique viral polypeptides-L, HN, Fo (F), NP, M, and a 47,000-dalton polypeptide. The small virion glycopolypeptide F is related to Fo, a glycopolypeptide found only in infected cells. In addition, several smaller polypeptides, including a 53,000dalton polypeptide found both in purified virions and in infected cells, are related to the nucleocaspid protein. Kinetic analysis of each viral polypeptide reveals that all of the major viral polypeptides, with the possible exception of L, are stable after an amino acid chase. A precursor-product relationship between Fo and F was not demonstrable by pulse-chase experiments. Also, almost the same relative amount of F, the putative product, was present in infected cultures after either 5 or 30 min of radioisotopic labeling. These results suggest that Fo is processed rapidly.

Journal of Virology, Apr 1, 1977

We have isolated 1S RNA from cytoplasmic extracts of Newcastle disease virus-infected Chinese ham... more We have isolated 1S RNA from cytoplasmic extracts of Newcastle disease virus-infected Chinese hamster ovary cells and tested its ability to direct protein synthesis in extracts derived from wheat germ. The products of the cell-free reaction directed by this RNA contain polypeptides that comigrate with NP, M, F, and 47K proteins from virions. In addition, the products contain a polypeptide (67K) that migrates on polyacrylamide gels slightly faster than the HN protein from virions. Tryptic peptide analysis of the cell-free products and proteins from virions confirms their identity.

Journal of Virology, Oct 1, 1978

The role of glycosylation in the maturation of the vesicular stomatitis virus (VSV) glycoprotein ... more The role of glycosylation in the maturation of the vesicular stomatitis virus (VSV) glycoprotein was studied by use of the antibiotic tunicamycin. Tunicamycin-treated VSV-infected cells synthesize an unglycosylated form of the VSV glycoprotein (R. Leavitt, S. Schlesinger, and S. Kornfeld, J. Virol. 21:375-385, 1977). We have found that tunicamycin has no effect on the attachment of the glycoprotein to intracellular membranes or on the transport of protein to the lumen of the endoplasmic reticulum. However, tunicamycin prevented the migration of the glycoprotein from the rough endoplasmic reticulum to smooth intracellular membranes.

Virus Research, May 1, 1987

Journal of Biological Chemistry, Sep 1, 1975

bioRxiv (Cold Spring Harbor Laboratory), Jul 30, 2021

Maternal anti-respiratory syncytial virus (RSV) antibodies acquired by the fetus through the plac... more Maternal anti-respiratory syncytial virus (RSV) antibodies acquired by the fetus through the placenta protect neonates from RSV disease through the first weeks of life. In the cotton rat model of RSV infections, we previously reported that immunization of dams during pregnancy with virus-like particles assembled with mutation stabilized pre-fusion F protein as well as the wild type G protein resulted in robust protection of their offspring from RSV challenge (Blanco,

Virus Research, Sep 1, 1986

Wiley Encyclopedia of Molecular Medicine, Jan 15, 2002

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Virus Research, Mar 1, 1995

Comparisons of the sequences of the hemagglutinin-neuraminidase (HN) protein from thirteen differ... more Comparisons of the sequences of the hemagglutinin-neuraminidase (HN) protein from thirteen different strains of Newcastle disease virus (NDV) show that while 12 cysteine residues are conserved in all strains, two cysteine residues are variably present (Sakaguchi et al. (1989) Virology 169, 260-272). One of these residues, at amino acid 6, is in the cytoplasmic domain. The other cysteine is at amino acid 123 in the ectodomain and is responsible for disulfide-linked HN dimers detected in some NDV strains (McGinnes and Morrison (1994) Virology 200, 470-483). To explore the role of these nonconserved residues in the structure and function of the protein, cysteine residues at amino acid 6 and 123 in the HN protein of the AV strain of NDV were mutated individually and in combination by site specific mutagenesis to serine and tryptophan, respectively. Proteins with mutations in either residue (C6S or C123W) or in both residues (C6S,123W) were transported to the cell surface. However, all three mutants had reduced attachment, neuraminidase, and fusion promotion activities. All three mutant proteins also showed an alteration in an antigenic site specific for oligomers of HN protein while all other antigenic sites were present at wild type levels. These results suggest that the nonconserved cysteine residues in the HN sequence may modulate the biological activities of the protein by affecting the oligomeric structure of the protein.

Journal of Virology, Feb 1, 1977

We report here an in vitro system designed to study the interactions of vesicular stomatitis viru... more We report here an in vitro system designed to study the interactions of vesicular stomatitis virus (VSV) proteins with cellular membranes. We have synthesized the VSV nucleocapsid (N) protein, nonstructural (NS) protein, glycoprotein (G protein), and membrane (M) protein in a wheat germ, cell-free, protein-synthesizing system directed by VSV 12 to 18S RNA. When incubated at low salt concentrations with purified cytoplasmic membranes derived from Chinese hamster ovary cells, the VSV M and G proteins bind to membranes, whereas the VSV N and NS proteins do not. The VSV M protein binds to membranes in low or high divalent cation concentrations, whereas binding of significant amounts of G protein requires at least 5 mM magnesium acetate concentrations. Vesicular stomatitis virus (VSV) is a simple, lar membranes, whereas the VSV N and NS enveloped virus that contains two membrane proteins do not. proteins: the glycoprotein (G protein), which forms the spikes of the virion (4, 23), and the MATERIALS AND METHODS membrane (M) protein, which lines the inner Cells and viruses. Membranes were prepared surface of the viral membrane (3). There are from CHO cells. Stocks of VSV (pure B particles of three other known viral proteins, the VSV nuthe Indiana serotype) were grown in CHO cells and cleocapsid (N) protein, the nonstructural (NS) purified as described previously (20). protein, and the viral transcriptase (L) protein. Preparation of VSV 12-186 polyribosomal RNA. These three proteins are associated with the The procedure described by Palmiter (15) was used core ofthe virus particle (16, 23, 24). Each of the with several modifications for the preparation of five viral proteins is synthesized from a mono-VSV 12-18S polyribosomal RNA. CHO cells growing cistronic mRNA (9, 13, 14). at 37°C were infected with VSV at a multiplicity of 3 risngtic m arNAl(9 13, 14). PFU/cell as described previously (20), except that 5 During the early stages in the maturation Of~.tg of actinomycin D per ml was added at the beginthis virus, host cell membranes are modified ning of infection. [3H]uridine (70 Ci/mmol, 25 ,uCi/ with the VSV G and M proteins. Cell fractionaml; New England Nuclear Corp.) was added 2 h tion studies of VSV-infected cells have shown postinfection. Infected cells were harvested at 4.5 h that the VSV M and G proteins rapidly become postinfection, suspended in sucrose-TKM buffer associated with the membrane fraction of the (0.05 M Tris [pH 7.5], 0.025 M KCl, 0.005 M magnecells after their synthesis (5, 11, 12, 24). Nucleo-sium acetate, 0.25 M sucrose), and disrupted with 10 capsid structures containing the genome RNA strokes of a tight-fitting Dounce homogenizer. Nuas well as the viral N, NS, and L proteins are clei were removed by centrifugation (1,000 x g for 2 asswemlld as the vira plaNsm. andsLptroteisures min). The resulting cytoplasmic extract was centriassembled in the cytoplasm. These structures fuged at 20,000 x g for 20 min, and the pellet (mem

Journal of Virology, Mar 1, 1979

Journal of Virology, Jul 1, 1984

Upon infection, the Newcastle disease virus (NDV) genome is transcribed to produce 18S, 22S, and ... more Upon infection, the Newcastle disease virus (NDV) genome is transcribed to produce 18S, 22S, and 35S RNAs (M. Bratt, and W. Robinson, J. Mol. Biol. 23:1-21, 1967). The 22S RNA has been shown to contain 18S sequences and is thought to represent polycistronic transcripts generated by transcriptional readthrough of adjacent genes (Varich et al., Acta Virol. 23:341-343, 1979). With improved extraction procedures, the 22S RNA was found to represent up to 25 % of the total transcription in NDV-infected cells. This RNA was resolved into at least five discrete species on formaldehyde-agarose gels. All but one of these molecules contain 3' polyadenylate sequences but not internal polyadenylate sequences. These transcripts are found on polyribosomes of infected cells, suggesting that they are functional mRNAs.

Journal of Virology, Oct 1, 1980

Polypeptides synthesized in Newcastle disease virus (NDV)-infected CHO cells in the absence of gl... more Polypeptides synthesized in Newcastle disease virus (NDV)-infected CHO cells in the absence of glycosylation were characterized. Incorporation of either [3H]mannose or [3H]glucosamine into NDV polypeptides was inhibited to greater than 99% by the antibiotic tunicamycin. Under these conditions, infected cells synthesized proteins which comigrated on polyacrylamide gels with the viral L protein, nucleocapsid protein, membrane protein, and a polypeptide with a molecular weight of 55,000 (P55). These cells did not synthesize polypeptides with the size of the hemagglutinin-neuraminidase (HN) protein or the fusion (Fo) protein. They did, however, synthesize new polypeptides with molecular weights of 75,000 (P75), 67,000 (P67), and 52,000 (P52). Peptide analysis revealed that P75 was a host cell protein whose synthesis is enhanced by tunicamycin. P67 corresponded to the unglycosylated HN protein, and P52 corresponded to a cleaved, unglycosylated form of Fo. These unglycosylated forms of the glycoproteins were found to be relatively stable in infected cells. P55, previously thought to correspond to the cleaved form of Fo, was found to be a unique viral protein which is associated with intracellular nucleocapsid structures.

Virus Research, Jul 1, 1986

During infection, the Newcastle disease virus (NDV) genome is transcribed to produce 5 to 7 speci... more During infection, the Newcastle disease virus (NDV) genome is transcribed to produce 5 to 7 species of polycistronic messenger RNA (Wilde and Morrison, J. Virol. 51, 71-76) in addition to the well characterized monocistronic messenger RNA. To identify the specific sequences present in each of the polycistronic RNA species, cDNA clones generated by reverse transcription of NDV mRNAs were characterized and used as probes on Northern blots of total NDV cytoplasmic RNA. By this method, it was shown that four of these large RNA species are polycistronic transcripts containing sequences from two genes: one species contains nucfeocapsid protein (NP) and phosphoprotein (P) gene sequences; another, P and membrane protein (M) gene sequences; another, M and fusion protein (F,) gene sequences; and another, FO and hema~utinin-neur~~dase protein (I-IN) gene sequences. The existence of these transcripts yields a transc~pti~n map order of NP, P, M, F,, HN. The remaining RNA bands may be composed of at least three different polycistronic transcripts, each of which represents transcription through three adjacent genes. Newcastle disease virus, transcription, polycistronic transcripts Intiuctlou Newcastle disease virus (NDV is a member of the p~arn~o~s group of viruses whose nonrated genome of 5100 to 5700 kDa (Kolakofsky et al., 1974) encodes the six known viral proteins (Bratt and Hightower, 1977

Virus Research, Feb 1, 1990

Characterization of the posttranslational modifications of the mature, cell-associated hemaggluti... more Characterization of the posttranslational modifications of the mature, cell-associated hemagglutinin-neur~~dase (HN) protein of Newcastle disease virus (NDV) revealed that the HN protein exists in two forms differentiated by disulfide bonds and glycosylation. One form, HN,, contains intermolecular disulfide bonds and is endoglycosidase H partially resistant. The other form, HN,, is not linked by disulfide bonds and is endoglycosidase H sensitive. Both forms of the protein are modified with fucose indicating transport to the Golgi membranes. Both forms are detected at the cell surface by monoclonal antibody. Furthermore, both forms are transported to the cell surface with identical kinetics. HN, is incorporated into virions. HN, is not incorporated into virions and is presumably degraded. The cDNA derived from the HN gene was expressed from a retrovirus vector. The majority of the protein expressed was in the nonvi~on-ass~iated form b. Evidence is presented that the level of gene expression determines the ratio of the two forms of HN protein. At high levels of expression, the virion-associated form is favored while at low levels of expression the nonvirion-associated form is favored. The results presented have implications for persistent infections as well as expression of viral genes from different vectors. Newcastle disease virus; HN protein

Human Vaccines & Immunotherapeutics, Dec 12, 2022

Journal of Virology, Dec 1, 2019

Maternal vaccination may be the most effective and safest approach to the protection of infants f... more Maternal vaccination may be the most effective and safest approach to the protection of infants from respiratory syncytial virus (RSV) infection, a severe acute lower respiratory tract disease in infants and young children worldwide. We previously compared five different virus-like particle (VLP)-associated, mutationstabilized prefusion F (pre-F) proteins, including the prototype DS-Cav1 F VLPs. We showed that alternative versions of prefusion F proteins have different conformations and induce different populations of anti-F protein antibodies. Two of these alternative pre-F VLPs, the UC-2 F and UC-3 F VLPs, stimulated in mice higher titers of neutralizing antibodies than DS-Cav1 F VLPs (M.

Journal of Virology, Mar 1, 1985

The fusion glycoprotein (Fo) of Newcastle disease virus is cleaved at an intracellular site (Naga... more The fusion glycoprotein (Fo) of Newcastle disease virus is cleaved at an intracellular site (Nagai et al., Virology 69:523-538, 1976) into F1 and F2. This result was confirmed by comparing the transit time of the fusion protein to the cell surface with the time course of cleavage of Fo. The time required for cleavage of half of the pulse-labeled Fo protein is ca. 40 min faster than the half time of the transit of the fusion protein to the cell surface. To determine the cell compartment in which cleavage occurs, use was made of inhibitors which block glycoprotein migration at specific points and posttranslational modifications known to occur in specific cell membranes. Cleavage of Fo is inhibited by carbonyl cyanide m-chlorophenylhydrazone; thus, cleavage does not occur in the rough endoplasmic reticulum. Monensin blocks the incorporation of Newcastle disease virus glycoproteins into virions and blocks the cleavage of the fusion glycoprotein. However, Fo cannot be radioactively labeled with [3H] fucose, whereas F1 is readily labeled. These results argue that cleavage occurs in the trans Golgi membranes or in a cell compartment occupied by glycoproteins quite soon after their transit through the trans Golgi membranes. The implications of the results presented for the transit times of the fusion protein between subcellular organelles are discussed.

Journal of Virology

Respiratory syncytial virus (RSV) results in significant disease in infants, young children, and ... more Respiratory syncytial virus (RSV) results in significant disease in infants, young children, and the elderly. Thus, development of an effective vaccine for these populations is a priority.