bluuebunny/crossref_metadata_2025_split huggingface dataset:
Interaction with the p6 Domain of the Gag Precursor Mediates Incorporation into Virions of Vpr and Vpx Proteins from Primate Lentiviruses
Authors: Selig L. , Pages J.-C. , Tanchou V. , Prévéral S. , Berlioz-Torrent C. , Liu L. X. , Erdtmann L. , Darlix J.-L. , Benarous R. , Benichou S. 
Published: December 2019

ABSTRACT

Vpr and Vpx proteins from human and simian immunodeficiency viruses (HIV and SIV) are incorporated into virions in quantities equivalent to those of the viral Gag proteins. We demonstrate here that Vpr and Vpx proteins from distinct lineages of primate lentiviruses were able to bind to their respective Gag precursors. The capacity of HIV type 1 (HIV-1) Vpr mutants to bind to Pr55
Gag
was correlated with their incorporation into virions. Molecular analysis of these interactions revealed that they required the C-terminal p6 domain of the Gag precursors. While the signal for HIV-1 Vpr binding lies in the leucine triplet repeat region of the p6 domain reported to be essential for incorporation, SIV
sm
Gag lacking the equivalent region still bound to SIV
sm
Vpr and Vpx, indicating that the determinants for Gag binding are located upstream of this region of the p6 domain. Binding to Gag cleavage products showed that HIV-1 Vpr interacted directly with the nucleocapsid protein (NC), whereas SIV
sm
Vpr and Vpx did not interact with NC but with the p6 protein. These results (i) reveal differences between HIV-1 and SIV
sm
for the p6 determinants required for Vpr and Vpx binding to Gag and (ii) suggest that HIV-1 Vpr and SIV
sm
Vpr and Vpx interact with distinct cleavage products of the precursor following proteolytic processing in the virions.
Herpes Simplex Virus Type 1 Immediate-Early Protein Vmw110 Induces the Proteasome-Dependent Degradation of the Catalytic Subunit of DNA-Dependent Protein Kinase
Authors: Parkinson Jane , Lees-Miller Susan P. , Everett Roger D. 
Published: December 2019

ABSTRACT
Herpes simplex virus type 1 (HSV-1) infection causes the active degradation of the catalytic subunit of DNA-dependent protein kinase (DNA-PKcs), and this process is reliant on the expression of the HSV-1 immediate-early protein Vmw110. In this study we investigated in more detail the mechanism by which the degradation occurs, the domains of Vmw110 which are required, and whether Vmw110 is by itself sufficient for the effect. We found that proteasome inhibitors prevented the degradation of DNA-PKcs, indicating the involvement of a proteasome pathway. Furthermore, the continued activity of DNA-PK during infection in the presence of these inhibitors indicated that Vmw110 does not directly alter the enzyme activity of DNA-PKcs prior to its degradation in a normal infection. Indeed, Vmw110 was found to bind to neither the catalytic nor Ku subunits of DNA-PK. Using mutant Vmw110 viruses we show that the RING finger domain of Vmw110 is essential for the induced degradation of DNA-PKcs but that the ability of Vmw110 to bind to a cellular ubiquitin-specific protease (HAUSP) is not required. When expressed in the absence of other viral proteins, Vmw110 was sufficient to cause the degradation of DNA-PKcs, indicating that the effect on the stability of DNA-PKcs was a direct consequence of Vmw110 activity and not an indirect Vmw110-dependent effect of virus infection. Finally, the Vmw110-induced degradation of DNA-PKcs and loss in DNA-PK activity appears to be beneficial to HSV-1 infection, as virus replication was more efficient in cells lacking DNA-PKcs, especially at low multiplicities of infection.
Antigen-Specific Expansion of Cytotoxic T Lymphocytes in Acute Measles Virus Infection
Authors: Mongkolsapaya Juthathip , Jaye Assan , Callan Margaret F. C. , Magnusen Albert F. , McMichael Andrew J. , Whittle Hilton C. 
Published: December 2019

ABSTRACTSkewing of the T-cell receptor repertoire of CD8+T cells has been shown in some persistent infections with viruses, such as human immunodeficiency virus, simian immunodeficiency virus, and Epstein-Barr virus. We have demonstrated that similar distortions also occur in nonpersistent measles virus infection. In addition, two of four children immunized with live, attenuated measles virus showed larger and more persistent CD8+T-cell expansions than their naturally infected counterparts. The expanded lymphocyte populations were monoclonal or oligoclonal and lysed target cells infected with recombinant vaccinia virus expressing measles virus protein. These results demonstrate that the expansions of CD8+T lymphocytes are antigen driven.
Polypyrimidine Tract-Binding Protein Binds to the Leader RNA of Mouse Hepatitis Virus and Serves as a Regulator of Viral Transcription
Authors: Li Hsin-Pai , Huang Peiyong , Park Sungmin , Lai Michael M. C. 
Published: December 2019

ABSTRACT
A cellular protein, previously described as p55, binds specifically to the plus strand of the mouse hepatitis virus (MHV) leader RNA. We have purified this protein and determined by partial peptide sequencing that it is polypyrimidine tract-binding protein (PTB) (also known as heterogeneous nuclear ribonucleoprotein [hnRNP] I), a nuclear protein which shuttles between the nucleus and cytoplasm. PTB plays a role in the regulation of alternative splicing of pre-mRNAs in normal cells and translation of several viruses. By UV cross-linking and immunoprecipitation studies using cellular extracts and a recombinant PTB, we have established that PTB binds to the MHV plus-strand leader RNA specifically. Deletion analyses of the leader RNA mapped the PTB-binding site to the UCUAA pentanucleotide repeats. Using a defective-interfering RNA reporter system, we have further shown that the PTB-binding site in the leader RNA is critical for MHV RNA synthesis. This and our previous study (H.-P. Li, X. Zhang, R. Duncan, L. Comai, and M. M. C. Lai, Proc. Natl. Acad. Sci. USA 94:9544–9549, 1997) combined thus show that two cellular hnRNPs, PTB and hnRNP A1, bind to the transcription-regulatory sequences of MHV RNA and may participate in its transcription.
Characterization of a Natural Mutation in an Antigenic Site on the Fusion Protein of Measles Virus That Is Involved in Neutralization
Authors: Fayolle Joël , Verrier Bernard , Buckland Robin , Wild T. Fabian 
Published: December 2019

ABSTRACT
Although measles virus is an antigenically monotypic virus, nucleotide sequence analysis of the hemagglutinin and nucleoprotein genes has permitted the differentiation of a number of genotypes. In contrast, the fusion (F) protein is highly conserved; only three amino acid changes have been reported over a 40-year period. We have isolated a measles virus strain which did not react with an anti-F monoclonal antibody (MAb) which we had previously shown to be directed against a dominant antigenic site. This virus strain, Lys-1, had seven amino acid changes compared with the Edmonston strain. We have shown that a single amino acid at position 73 is responsible for its nonreactivity with the anti-F MAb. With the same MAb, antibody-resistant mutants were prepared from the vaccine strain. A single amino acid change at position 73 (R→W) was observed. The possibility of selecting measles virus variants in vaccinated populations is discussed.
An Attenuated Variant of the GDVII Strain of Theiler’s Virus Does Not Persist and Does Not Infect the White Matter of the Central Nervous System
Authors: Jarousse Nadine , Viktorova Ekaterina G. , Pilipenko Evgeny V. , Agol Vadim I. , Brahic Michel 
Published: December 2019

ABSTRACT
The DA strain of Theiler’s virus causes a persistent and demyelinating infection of the white matter of spinal cord, whereas the GDVII strain causes a fatal gray-matter encephalomyelitis. Studies with recombinant viruses showed that this difference in phenotype is controlled mainly by the capsid. However, conflicting results regarding the existence of determinants of persistence in the capsid of the GDVII strain have been published. Here we show that a GDVII virus whose neurovirulence has been attenuated by an insertion in the 5′ noncoding region does not persist in the central nervous systems of mice. Furthermore, this virus infects the gray matter efficiently, but not the white matter. These results confirm the absence of determinants of persistence in the GDVII capsid. They suggest that the DA capsid controls persistence by allowing the virus to infect cells in the white matter of the spinal cord.
Transcriptional Activation Signals Found in the Epstein-Barr Virus (EBV) Latency C Promoter Are Conserved in the Latency C Promoter Sequences from Baboon and Rhesus Monkey EBV-Like Lymphocryptoviruses (Cercopithicine Herpesviruses 12 and 15)
Authors: Fuentes-Pananá Ezequiel M. , Swaminathan Sankar , Ling Paul D. 
Published: December 2019

ABSTRACT
The Epstein-Barr virus (EBV) EBNA2 protein is a transcriptional activator that controls viral latent gene expression and is essential for EBV-driven B-cell immortalization. EBNA2 is expressed from the viral C promoter (Cp) and regulates its own expression by activating Cp through interaction with the cellular DNA binding protein CBF1. Through regulation of Cp and EBNA2 expression, EBV controls the pattern of latent protein expression and the type of latency established. To gain further insight into the important regulatory elements that modulate Cp usage, we isolated and sequenced the Cp regions corresponding to nucleotides 10251 to 11479 of the EBV genome (−1079 to +144 relative to the transcription initiation site) from the EBV-like lymphocryptoviruses found in baboons (herpesvirus papio; HVP) and Rhesus macaques (RhEBV). Sequence comparison of the approximately 1,230-bp Cp regions from these primate viruses revealed that EBV and HVP Cp sequences are 64% conserved, EBV and RhEBV Cp sequences are 66% conserved, and HVP and RhEBV Cp sequences are 65% conserved relative to each other. Approximately 50% of the residues are conserved among all three sequences, yet all three viruses have retained response elements for glucocorticoids, two positionally conserved CCAAT boxes, and positionally conserved TATA boxes. The putative EBNA2 100-bp enhancers within these promoters contain 54 conserved residues, and the binding sites for CBF1 and CBF2 are well conserved. Cp usage in the HVP- and RhEBV-transformed cell lines was detected by S1 nuclease protection analysis. Transient-transfection analysis showed that promoters of both HVP and RhEBV are responsive to EBNA2 and that they bind CBF1 and CBF2 in gel mobility shift assays. These results suggest that similar mechanisms for regulation of latent gene expression are conserved among the EBV-related lymphocryptoviruses found in nonhuman primates.
Feline Calicivirus Capsid Protein Expression and Capsid Assembly in Cultured Feline Cells
Authors: Geissler Klaus , Schneider Karla , Fleuchaus Andrea , Parrish Colin R. , Sutter Gerd , Truyen Uwe 
Published: December 2019

ABSTRACT
The capsid protein of feline calicivirus (FCV) was expressed by using plasmids containing cytomegalovirus, simian virus 40, or T7 promoters. The strongest expression was achieved with the T7 promoter and coinfection with vaccinia virus expressing the T7 RNA polymerase (MVA/T7pol). The FCV precursor capsid protein was processed to the mature-size protein, and these proteins were assembled in to virus-like particles.
Binding Specificity of Protein Phosphatase 2A Core Enzyme for Regulatory B Subunits and T Antigens
Authors: Ruediger Ralf , Fields Katherine , Walter Gernot 
Published: December 2019

ABSTRACT
The core enzyme of protein phosphatase 2A is composed of a regulatory subunit A and a catalytic subunit C. It is controlled by three types of regulatory B subunits (B, B′, and B") and by tumor (T) antigens, which are unrelated by sequence but bind to overlapping regions on the A subunit. To find out whether the different B subunits and T antigens bind to identical or distinct amino acids of the A subunit, mutants were generated and their abilities to bind B subunits and T antigens were tested. We found that some amino acids are involved in the binding of all types of B subunits, whereas others are specifically involved in the binding of one or two types of B subunits. T-antigen-binding specificity does not correlate with that of a particular type of B subunit.
Constrained Evolution of Human Immunodeficiency Virus Type 1 Protease during Sequential Therapy with Two Distinct Protease Inhibitors
Authors: Dulioust Anne , Paulous Sylvie , Guillemot Laurent , Delavalle Anne-Marie , Boué François , Clavel François 
Published: December 2019

ABSTRACT
Human immunodeficiency virus type 1 (HIV-1) variants that have developed protease (PR) inhibitor resistance most often display cross-resistance to several molecules within this class of antiretroviral agents. The clinical benefit of the switch to a second PR inhibitor in the presence of such resistant viruses may be questionable. We have examined the evolution of HIV-1 PR genotypes and phenotypes in individuals having failed sequential treatment with two distinct PR inhibitors: saquinavir (SQV) followed by indinavir (IDV). In viruses where typical SQV resistance mutations were detected before the change to IDV, the corresponding mutations were maintained under IDV, while few additional mutations emerged. In viruses where no SQV resistance mutations were detected before the switch to IDV, typical SQV resistance profiles emerged following the introduction of IDV. We conclude that following suboptimal exposure to a first PR inhibitor, the introduction of a second molecule of this class can lead to rapid selection of cross-resistant virus variants that may not be detectable by current genotyping methods at the time of the inhibitor switch. Viruses committed to resistance to the first inhibitor appear to bear the “imprint” of this initial selection and can further adapt to the selective pressure exerted by the second inhibitor following a pathway that preserves most of the initially selected mutations.
Rapid Clearance of Simian Immunodeficiency Virus Particles from Plasma of Rhesus Macaques
Authors: Zhang Linqi , Dailey Peter J. , He Tian , Gettie Agegnehu , Bonhoeffer Sebastian , Perelson Alan S. , Ho David D. 
Published: December 2019

ABSTRACTPerturbation of the equilibrium between human immunodeficiency virus type 1 (HIV-1) and the infected host by administering antiretroviral agents has revealed the rapid turnover of both viral particles and productively infected cells. In this study, we used the infusion of simian immunodeficiency virus (SIV) particles into rhesus macaques to obtain a more accurate estimate of viral clearance in vivo. Consistently, exogenously infused virions were cleared from plasma with an extremely short half-life, on the order of minutes (a mean of 3.3 min). This new estimate is ∼100-fold lower than the upper bound of 6 h previously reported for HIV-1 in infected humans. In select animals, multiple tissues were collected at the completion of each experiment to track the potential sites of virion clearance. Detectable levels of SIV RNA were found in lymph nodes, spleen, lungs, and liver, but not in other tissues examined. However, only ∼1 to 10% or less of the infused virions were accounted for by the thorough tissue sampling, indicating that the vast majority of the infused particles must have been degraded over a short period of time. Should the rapid clearance of virions described here be applicable to infected patients, then HIV-1 production and thus the number of productively infected CD4+T lymphocytes or the viral burst size must be proportionally higher than previous minimal estimates.
Repression of Epstein-Barr Virus EBNA-1 Gene Transcription by pRb during Restricted Latency
Authors: Ruf Ingrid K. , Sample Jeffery 
Published: December 2019

ABSTRACT

During the restricted programs of Epstein-Barr virus (EBV) latency in EBV-associated tumors and a subpopulation of latently infected B cells in healthy EBV carriers, transcription of the EBV nuclear antigen 1 (EBNA-1) gene is mediated by the promoter Qp. Previously, two noncanonical E2F binding sites were identified within Qp. The role of E2F in the regulation of Qp, however, has been controversial and is undefined. Here we demonstrate that an E2F factor(s) within Burkitt lymphoma (BL) cells binds to a G/C-rich element [GGCG(C/G)] within the previously identified binding sites in Qp and prototypical E2F response elements. Furthermore, Qp-driven reporter gene expression could be efficiently repressed through either E2F binding site by the tumor suppressor pRb, a potent transcriptional repressor targeted to promoters during G
0
and the early G
1
phase of the cell cycle via its interaction with E2F; a mutant pRb (pRb
706
) lacking E2F binding capability was unable to repress Qp. However, we did not observe cell cycle variation in the expression of either EBNA-1 mRNA or protein in exponentially growing BL cells, consistent with previous predictions that Qp is constitutively active in these cells and with the extremely long
t
1/2
of EBNA-1. By contrast, within G
0
/G
1
in growth-arrested BL cells, EBNA-1 mRNA levels were twofold lower than in S phase, similar to the two- to eightfold differences in cell cycle expression of some cyclin mRNAs. Thus, although regulation of Qp is coupled to the cell cycle, this clearly has no impact on the level of EBNA-1 expressed in proliferating cells. We conclude, therefore, that the most important contribution of E2F to the regulation of Qp is to direct the pRb-mediated suppression of EBNA-1 expression within resting B cells, the principal reservoir of latent EBV. This would provide a means to restrict unneeded and potentially deleterious expression of EBNA-1 in a nonproliferating cell and to coordinate the activation of EBNA-1 expression necessary for EBV genome replication and maintenance upon reentry of the cell cycle in response to proliferative signals.
Structural Maturation of the Transmissible Gastroenteritis Coronavirus
Authors: Salanueva Iñigo J. , Carrascosa José L. , Risco Cristina 
Published: December 2019

ABSTRACT

During the life cycle of the transmissible gastroenteritis coronavirus (TGEV), two types of virus-related particles are detected in infected swine testis cells: large annular viruses and small dense viruses. We have studied the relationships between these two types of particles. Immunoelectron microscopy showed that they are closely related, since both large and small particles reacted equally with polyclonal and monoclonal antibodies specific for TGEV proteins. Monensin, a drug that selectively affects the Golgi complex, caused an accumulation of large annular viral particles in perinuclear elements of the endoplasmic reticulum-Golgi intermediate compartment. A partial reversion of the monensin blockade was obtained in both the absence and presence of cycloheximide, a drug that prevented the formation of new viral particles. After removal of monensin, the Golgi complex recovered its perinuclear location, and a decrease in the number of perinuclear large viral particles was observed. The release of small dense viral particles into secretory vesicles and the extracellular medium was also observed, as was a partial recovery of infectivity in culture supernatants. Small viral particles started to be seen between the third and the fourth Golgi cisternae of normally infected cells. All of these data strongly indicate that the large annular particles are the immature precursors of the small dense viruses, which are the infectious TGEV virions. The immature viral particles need to reach a particular location at the
trans
side of the Golgi stack to complete their morphological maturation.
Induction of Bcl-xLExpression by Human T-Cell Leukemia Virus Type 1 Tax through NF-κB in Apoptosis-Resistant T-Cell Transfectants with Tax
Authors: Tsukahara Tomonori , Kannagi Mari , Ohashi Takashi , Kato Hirotomo , Arai Masaaki , Nunez Gabriel , Iwanaga Youichi , Yamamoto Naoki , Ohtani Kiyoshi , Nakamura Masataka , Fujii Masahiro 
Published: December 2019

ABSTRACTHuman T-cell leukemia virus type 1 (HTLV-1) Tax is thought to play a pivotal role in immortalization of T cells. We have recently shown that the expression of Tax protected the mouse T-cell line CTLL-2 against apoptosis induced by interleukin-2 (IL-2) deprivation and converted its growth from being IL-2 dependent to being IL-2 independent. In this study, we demonstrate that constitutive expression ofbcl-xlbut notbcl-2,bcl-xs,bak,bad, orbaxwas associated with apoptosis resistance after IL-2 deprivation in CTLL-2 cells that expressed Tax. Transient-transfection assays showed thatbcl-xpromoter was transactivated by wild-type Tax. Similar effects were observed in mutant Tax retaining transactivating ability through NF-κB. Deletion or substitution of a putative NF-κB binding site identified in thebcl-xpromoter significantly decreased Tax-induced transactivation. This NF-κB-like element was able to form a complex with NF-κB family proteins in vitro. Furthermore, Tax-induced transactivation of thebcl-xpromoter was also diminished by the mutant IκBα, which specifically inhibits NF-κB activity. Our findings suggest that constitutive expression of Bcl-xLinduced by Tax through the NF-κB pathway contributes to the inhibition of apoptosis in CTLL-2 cells after IL-2 deprivation.
Regulation of Closterovirus Gene Expression Examined by Insertion of a Self-Processing Reporter and by Northern Hybridization
Authors: Hagiwara Yuka , Peremyslov Valery V. , Dolja Valerian V. 
Published: December 2019

ABSTRACT
A reporter open reading frame (ORF) coding for a fusion of bacterial β-glucuronidase (GUS) with a proteinase domain (Pro) derived from tobacco etch potyvirus was utilized for tagging individual genes of beet yellows closterovirus (BYV). Insertion of this reporter ORF between the first and second codons of the BYV ORFs encoding the HSP70 homolog (HSP70h), a major capsid protein (CP), and a 20-kDa protein (p20) resulted in the expression of the processed GUS-Pro reporter from corresponding subgenomic RNAs. The high sensitivity of GUS assays permitted temporal analysis of reporter accumulation, revealing early expression from the HSP70h promoter, followed by the CP promoter and later the p20 promoter. The kinetics of transcription of the remaining BYV genes encoding a 64-kDa protein (p64), a minor capsid protein (CPm), and a 21-kDa protein (p21) were examined via Northern blot analysis. Taken together, the data indicated that the temporal regulation of BYV gene expression includes early (HSP70h, CPm, CP, and p21 promoters) and late (p64 and p20 promoters) phases. It was also demonstrated that the deletion of six viral genes that are nonessential for RNA amplification resulted in a dramatic increase in the level of transcription from one of the two remaining subgenomic promoters. Comparison with other positive-strand RNA viruses producing multiple subgenomic RNAs showed the uniqueness of the pattern of closterovirus transcriptional regulation.
The Simian Immunodeficiency Virus Envelope Glycoprotein Contains Two Epitopes Presented by the Mamu-A*01 Class I Molecule
Authors: Furchner Michelle , Erickson Ann L. , Allen Todd , Watkins David I. , Sette Alessandro , Johnson Philip R. , Walker Christopher M. 
Published: December 2019

ABSTRACTCytotoxic T lymphocyte (CTL) responses against the simian immunodeficiency virus (SIV) envelope and Gag proteins were monitored in a Mamu-A*01-positive rhesus macaque infected with SIVsmE660. Peripheral blood mononuclear cells (PBMC) cultured with synthetic peptides spanning the entire gp160 and Gag coding region recognized a total of three epitopes. One located in Gag was identified as the previously described Mamu-A*01-restricted p11cC→M epitope (CTPYDINQM). The other two epitopes, designated p15m and p54m, were located in the gp160 envelope protein. Both were nine amino acids in length and were predicted to bind Mamu-A*01 because they contained proline and leucine residues at positions 3 and 9, respectively. Indeed, expression of this class I major histocompatibility complex molecule was required for target cell recognition by envelope-specific CD8+T cells directed against both epitopes. These Mamu-A*01-restricted epitopes in the SIV envelope will be useful for monitoring immune responses in vaccinated or infected animals.