Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 96
Filtrar
Mais filtros










Base de dados
Intervalo de ano de publicação
1.
J Virol ; 2020 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-32611749

RESUMO

Us3 proteins of herpes simplex virus 1 (HSV-1) and HSV-2 are multi-functional serine-threonine protein kinases. Here, we identified an HSV-2 tegument protein UL7 as a novel physiological substrate of HSV-2 Us3. Mutations in HSV-2 UL7, which precluded Us3 phosphorylation of the viral protein, significantly reduced mortality, viral replication in the vagina, and development of vaginal disease in mice following vaginal infection. These results indicated that Us3 phosphorylation of UL7 in HSV-2 was required for efficient viral replication and pathogenicity in vivo Of note, this phosphorylation was conserved in UL7 of chimpanzee herpesvirus (ChHV), which phylogenically forms a monophyletic group with HSV-2 and the resurrected last common ancestral UL7 for HSV-2 and ChHV. In contrast, the phosphorylation was not conserved in UL7 of HSV-1, which belongs to a sister clade of the monophyletic group, the resurrected last common ancestral UL7 for HSV-1, HSV-2, and ChHV, and UL7s of other members of the genus Simplexvirus that are phylogenically close to these viruses. Thus, evolution of Us3 phosphorylation of UL7 coincided with the phylogeny of simplexviruses. Furthermore, artificially induced Us3 phosphorylation of UL7 in HSV-1 had no effect on viral replication and pathogenicity in mice in contrast to phosphorylation in HSV-2. Our results suggest that HSV-2 and ChHV have acquired and maintained Us3 phospho-regulation of UL7 during their evolution because the phospho-regulation had an impact on viral fitness in vivo, whereas most other simplexviruses have not because the phosphorylation was not necessary for efficient fitness of the viruses in vivo IMPORTANCE It has been hypothesized that the evolution of protein phospho-regulation drives phenotypic diversity across species of organisms, which impacts fitness during their evolution. However, there is a lack of information regarding linkage between the evolution of viral phospho-regulation and phylogeny of virus species. In this study, we clarified the novel HSV-2 Us3 phospho-regulation of UL7 in infected cells, which is important for viral replication and pathogenicity in vivo We also showed that evolution of Us3 phospho-regulation of UL7 was linked to phylogeny of viruses that are phylogenically close to HSV-2 and to phosphorylation requirements for the efficient in vivo viral fitness of HSV-2 and HSV-1, which are representative of viruses that have or have not evolved phospho-regulation, respectively. This study reports the first evidence showing that evolution of viral phospho-regulation coincides with phylogeny of virus species and supports the hypothesis regarding the evolution of viral phospho-regulation during viral evolution.

2.
Intern Med ; 59(5): 649-656, 2020 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-31735797

RESUMO

Pancreatic serous cystic neoplasms (SCNs), such as serous cystadenoma (SCA), are generally recognized as benign because malignant counterparts of SCNs have been extremely rare. In clinical practice, pancreatic cystic neoplasms diagnosed as SCNs have been managed by conservative observation, as long as the patients remained asymptomatic. We herein report a case of metachronous ductal adenocarcinoma that was discovered during long-term follow-up of SCN and review the related literature. To our knowledge, this was the first reported case of the local presence of ductal adenocarcinoma adjacent to SCA that was preoperatively diagnosed by endoscopic ultrasound-guided fine-needle aspiration.

3.
PLoS One ; 14(8): e0221468, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31437218

RESUMO

BACKGROUND AND AIMS: The revised Atlanta classification is widely used for the evaluation of acute pancreatitis (AP) severity. However, this classification cannot be used within 48 hours of AP onset. The aim of this study was to investigate the predictive factors of mortality in patients with AP on admission. METHODS: We evaluated the association between AP mortality and clinical parameters at the time of admission in patients with AP from April 2013 to December 2017 at one university hospital and one tertiary care referral center. RESULTS: A total of 203 consecutive patients were enrolled. Nine patients (4.4%) died despite multidisciplinary treatment. In a multivariable analysis, hematocrit ≥ 40% (odds ratio [OR], 1.07; 95% confidence interval [CI], 1.01-1.13; P = 0.021), blood urea nitrogen (BUN) ≥ 40 mg/dL (OR, 1.26; 95% CI, 1.11-1.42; P < 0.001), base excess < -3.0 mmol/L (OR, 1.15; 95% CI, 1.04-1.26; P = 0.004), and inflammation extending to the rectovesical excavation (OR, 1.19; 95% CI, 1.10-1.30; P < 0.001) on admission were significantly associated with mortality. CONCLUSION: Among the imaging findings, inflammation extending to the rectovesical excavation was the only independent predictive factor for mortality in AP. This simple finding, obtained on computed tomography without contrast agent on admission, might be a promising prognostic factor for AP.


Assuntos
Hospitalização , Pancreatite/mortalidade , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Inflamação/complicações , Inflamação/patologia , Masculino , Pessoa de Meia-Idade , Pancreatite/complicações , Pancreatite/diagnóstico por imagem , Pancreatite/economia , Valor Preditivo dos Testes , Índice de Gravidade de Doença , Resultado do Tratamento , Adulto Jovem
4.
J Virol ; 93(21)2019 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-31391274

RESUMO

During nuclear egress of nascent progeny herpesvirus nucleocapsids, the nucleocapsids acquire a primary envelope by budding through the inner nuclear membrane of infected cells into the perinuclear space between the inner and outer nuclear membranes. Herpes simplex virus 1 (HSV-1) UL34 and UL31 proteins form a nuclear egress complex (NEC) and play critical roles in this budding process, designated primary envelopment. To clarify the role of NEC binding to progeny nucleocapsids in HSV-1 primary envelopment, we established an assay system for HSV-1 NEC binding to nucleocapsids and capsid proteins in vitro Using this assay system, we showed that HSV-1 NEC bound to nucleocapsids and to capsid protein UL25 but not to the other capsid proteins tested (i.e., VP5, VP23, and UL17) and that HSV-1 NEC binding of nucleocapsids was mediated by the interaction of NEC with UL25. UL31 residues arginine-281 (R281) and aspartic acid-282 (D282) were required for efficient NEC binding to nucleocapsids and UL25. We also showed that alanine substitution of UL31 R281 and D282 reduced HSV-1 replication, caused aberrant accumulation of capsids in the nucleus, and induced an accumulation of empty vesicles that were similar in size and morphology to primary envelopes in the perinuclear space. These results suggested that NEC binding via UL31 R281 and D282 to nucleocapsids, and probably to UL25 in the nucleocapsids, has an important role in HSV-1 replication by promoting the incorporation of nucleocapsids into vesicles during primary envelopment.IMPORTANCE Binding of HSV-1 NEC to nucleocapsids has been thought to promote nucleocapsid budding at the inner nuclear membrane and subsequent incorporation of nucleocapsids into vesicles during nuclear egress of nucleocapsids. However, data to directly support this hypothesis have not been reported thus far. In this study, we have present data showing that two amino acids in the membrane-distal face of the HSV-1 NEC, which contains the putative capsid binding site based on the solved NEC structure, were in fact required for efficient NEC binding to nucleocapsids and for efficient incorporation of nucleocapsids into vesicles during primary envelopment. This is the first report showing direct linkage between NEC binding to nucleocapsids and an increase in nucleocapsid incorporation into vesicles during herpesvirus primary envelopment.

5.
Intern Med ; 58(21): 3087-3092, 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31292382

RESUMO

A 70-year-old man was admitted to our hospital due to elevated levels of hepatobiliary and pancreatic enzymes. Computed tomography showed contrast-enhanced mucosal hypertrophy from the duodenal papilla to the distal bile duct. Endoscopic examinations revealed a laterally spreading granular tumor and ampullary swelling. After surgical resection, an examination revealed well-differentiated adenocarcinoma of the ampulla with tubular adenoma spreading from the distal common bile duct to the second part of the duodenum showing both bile duct and duodenal phenotypes. To our knowledge, this is the first case of a tumor spreading from the bile duct to the duodenum that exhibited multiple phenotypes.


Assuntos
Adenocarcinoma/patologia , Adenoma/patologia , Neoplasias do Ducto Colédoco/patologia , Neoplasias Duodenais/patologia , Neoplasias Primárias Múltiplas/patologia , Adenocarcinoma/cirurgia , Adenoma/cirurgia , Idoso , Ampola Hepatopancreática , Ducto Colédoco/diagnóstico por imagem , Ducto Colédoco/patologia , Neoplasias do Ducto Colédoco/cirurgia , Neoplasias Duodenais/diagnóstico por imagem , Neoplasias Duodenais/cirurgia , Duodeno/diagnóstico por imagem , Duodeno/patologia , Humanos , Masculino , Invasividade Neoplásica , Neoplasias Primárias Múltiplas/cirurgia , Tomografia Computadorizada por Raios X
8.
J Virol ; 93(14)2019 07 15.
Artigo em Inglês | MEDLINE | ID: mdl-31043535

RESUMO

During the nuclear export of nascent nucleocapsids of herpes simplex virus 1 (HSV-1), the nucleocapsids acquire a primary envelope by budding through the inner nuclear membrane into the perinuclear space between the inner and outer nuclear membranes. This unique budding process, termed primary envelopment, is initiated by the nuclear egress complex (NEC), composed of the HSV-1 UL31 and UL34 proteins. Earlier biochemical approaches have shown that the NEC has an intrinsic ability to vesiculate membranes through the formation of a hexagonal lattice structure. The significance of intrahexamer interactions of the NEC in the primary envelopment of HSV-1-infected cells has been reported. In contrast, the contribution of lattice formation of the NEC hexamer to primary envelopment in HSV-1-infected cells remains to be elucidated. Therefore, we constructed and characterized a recombinant HSV-1 strain carrying an amino acid substitution in a UL31 residue that is an interhexamer contact site for the lattice formation of the NEC hexamer. This mutation was reported to destabilize the interhexamer interactions of the HSV-1 NEC. Here, we demonstrate that the mutation causes the aberrant accumulation of nucleocapsids in the nucleus and reduces viral replication in Vero and HeLa cells. Thus, the ability of HSV-1 to form the hexagonal lattice structure of the NEC was linked to an increase in primary envelopment and viral replication. Our results suggest that the lattice formation of the NEC hexamer has an important role in HSV-1 replication by regulating primary envelopment.IMPORTANCE The scaffolding proteins of several envelope viruses required for virion assembly form high-order lattice structures. However, information on the significance of their lattice formation in infected cells is limited. Herpesviruses acquire envelopes twice during their viral replication. The first envelop acquisition (primary envelopment) is one of the steps in the vesicle-mediated nucleocytoplasmic transport of nascent nucleocapsids, which is unique in biology. HSV-1 NEC, thought to be conserved in all members of the Herpesviridae family, is critical for primary envelopment and was shown to form a hexagonal lattice structure. Here, we investigated the significance of the interhexamer contact site for hexagonal lattice formation of the NEC in HSV-1-infected cells and present evidence suggesting that the lattice formation of the NEC hexamer has an important role in HSV-1 replication by regulating primary envelopment. Our results provide insights into the mechanisms of the envelopment of herpesviruses and other envelope viruses.

11.
Surg Endosc ; 33(12): 4026-4031, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-30759276

RESUMO

BACKGROUND AND AIMS: Migration of duodenal covered self-expandable metallic stents (C-SEMS) is the main cause of stent dysfunction in patients with malignant gastric outlet obstruction. However, the ideal method to prevent migration has not been clarified. We aimed to evaluate the feasibility and safety of duodenal C-SEMS fixation in this experimental study. METHODS: We used the over-the-scope clip (OTSC), suture, and clip methods to fix duodenal C-SEMS and evaluated the gripping force of each device and invasion depth based on pathological findings. RESULTS: The OTSC and suturing systems had a significantly higher mean gripping force compared with the clipping system (OTSC vs. clip: 13.2 vs. 1.0 Newtons [N], P < 0.001; suture vs. clip: 8.5 vs. 1.0 N, P < 0.001). OTSC compression was stronger compared with suturing (OTSC vs. suture: 13.2 vs. 8.5 N, P = 0.006). The submucosal layer, but not the muscle layer, was compressed more widely and deeply by OTSC compared with clips based on pathological findings by hematoxylin and eosin staining. CONCLUSION: Both OTSC and suturing methods used for duodenal C-SEMS fixation were feasible compared with the clipping method. The pathological evaluation of invasion depth indicated that OTSC may be safe even for preventive use. This study suggested that these methods can be applied clinically for duodenal C-SEMS fixation.

12.
Int Urol Nephrol ; 50(10): 1913-1918, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30182293

RESUMO

BACKGROUND: Reportedly, thiamine deficiency, resulting from malnutrition and long-term diuretic therapy, is observed in patients with chronic kidney disease (CKD). The risk of thiamine deficiency might be enhanced, especially in end-stage CKD patients. Here, we assessed thiamine status in incident dialysis patients. METHODS: This study was a single-center cross-sectional study which included 288 consecutive patients initiated into dialysis between April 2013 and March 2017 at our hospital. Thiamine status was evaluated by high-performance liquid chromatography of whole blood samples. We evaluated the association between blood thiamine concentration and other clinical parameters. RESULTS: Of the 288 patients, 21 patients receiving thiamine supplementation at the time of dialysis initiation and 26 patients without blood thiamine measurements were excluded. In 30 patients (12.4%), blood thiamine concentration was lower than the lower limit of normal (21.3 ng/mL; dotted line). Blood thiamine concentration correlated with age, body mass index, and Barthel index (BI) score (p = 0.008, 0.012 and 0.009, respectively). Stepwise multivariate regression analysis indicated that BI scores were independent risk factors for thiamine deficiency (ß coefficients = 0.169, p = 0.013). CONCLUSIONS: The proportion of end-stage CKD patients with low blood thiamine concentration is high. Low physical function (low BI score) is an independent risk factor of thiamine deficiency. Clinicians should be aware of thiamine deficiency in end-stage CKD patients, especially those with low physical function.


Assuntos
Falência Renal Crônica/sangue , Deficiência de Tiamina/sangue , Tiamina/sangue , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Índice de Massa Corporal , Estudos Transversais , Feminino , Nível de Saúde , Humanos , Falência Renal Crônica/complicações , Falência Renal Crônica/terapia , Masculino , Pessoa de Meia-Idade , Diálise Renal , Fatores de Risco , Deficiência de Tiamina/complicações , Deficiência de Tiamina/diagnóstico
13.
Dig Dis Sci ; 63(12): 3376-3381, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30206756

RESUMO

BACKGROUND: The ability of fluorescence in situ hybridization (FISH) assays in endoscopic transpapillary bile duct biopsy specimens to predict the prognosis of cholangiocarcinoma (CCA) has not been elucidated. AIMS: We aimed to clarify the association between the results of UroVysion FISH assays and the prognosis of CCA. METHODS: We retrospectively reviewed 49 specimens obtained by transpapillary forceps biopsy from consecutive patients with CCA. The copy numbers of chromosomes 3, 7, and 17 were evaluated by FISH assay using UroVysion. We compared the overall survival (OS) of CCA patients with and without increased copy numbers of chromosomes 3, 7, and 17. Furthermore, we evaluated the association between OS and the clinicopathological parameters of CCA patients. RESULTS: The OS was significantly shorter in patients with than without an increased chromosome 7 copy number (log-rank p = 0.015; median OS 11.9 vs. 20.7 months). In the univariate analyses, age (p = 0.012), ECOG performance status (p = 0.046), tumor stage (p = 0.046), surgery (p = 0.006), and an increased chromosome 7 copy number (p = 0.017) were significantly associated with OS. The multivariate analysis revealed that an increased chromosome 7 copy number (hazard ratio, 2.46; 95% CI 1.15-5.27; p = 0.021) and advanced clinical stage (hazard ratio, 2.26; 95% CI 1.11-4.63; p = 0.025) were independently predictive of a poor OS. CONCLUSIONS: Detection by FISH assay of an increased chromosome 7 copy number in transpapillary forceps biopsy specimens is predictive of a poor prognosis in CCA patients.


Assuntos
Neoplasias dos Ductos Biliares/genética , Neoplasias dos Ductos Biliares/patologia , Colangiocarcinoma/genética , Colangiocarcinoma/patologia , Cromossomos Humanos Par 7/genética , Dosagem de Genes/genética , Idoso , Idoso de 80 Anos ou mais , Neoplasias dos Ductos Biliares/mortalidade , Ductos Biliares Intra-Hepáticos/química , Ductos Biliares Intra-Hepáticos/patologia , Colangiocarcinoma/mortalidade , Feminino , Humanos , Hibridização in Situ Fluorescente/métodos , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Prognóstico , Estudos Retrospectivos , Taxa de Sobrevida/tendências
14.
Nat Immunol ; 19(10): 1071-1082, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-30201994

RESUMO

TLR3 is a sensor of double-stranded RNA that is indispensable for defense against infection with herpes simplex virus type 1 (HSV-1) in the brain. We found here that TLR3 was required for innate immune responses to HSV-1 in neurons and astrocytes. During infection with HSV-1, TLR3 recruited the metabolic checkpoint kinase complex mTORC2, which led to the induction of chemokines and trafficking of TLR3 to the cell periphery. Such trafficking enabled the activation of molecules (including mTORC1) required for the induction of type I interferons. Intracranial infection of mice with HSV-1 was exacerbated by impairment of TLR3 responses with an inhibitor of mTOR and was significantly 'rescued' by potentiation of TLR3 responses with an agonistic antibody to TLR3. These results suggest that the TLR3-mTORC2 axis might be a therapeutic target through which to combat herpes simplex encephalitis.


Assuntos
Encefalite por Herpes Simples/imunologia , Alvo Mecanístico do Complexo 2 de Rapamicina/imunologia , Receptor 3 Toll-Like/imunologia , Animais , Herpesvirus Humano 1 , Imunidade Inata/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Células NIH 3T3
16.
Nat Commun ; 9(1): 3379, 2018 08 23.
Artigo em Inglês | MEDLINE | ID: mdl-30139939

RESUMO

Vesicle-mediated nucleocytoplasmic transport is a nuclear pore-independent mechanism for the nuclear export of macromolecular complexes, but the molecular basis for this transport remains largely unknown. Here we show that endosomal sorting complex required for transport-III (ESCRT-III) is recruited to the inner nuclear membrane (INM) during the nuclear export of herpes simplex virus 1 (HSV-1). Scission during HSV-1 budding through the INM is prevented by depletion of ESCRT-III proteins. Interestingly, in uninfected human cells, the depletion of ESCRT-III proteins induces aberrant INM proliferation. Our results show that HSV-1 expropriates the ESCRT-III machinery in infected cells for scission of the INM to produce vesicles containing progeny virus nucleocapsids. In uninfected cells, ESCRT-III regulates INM integrity by downregulating excess INM.


Assuntos
Transporte Ativo do Núcleo Celular/fisiologia , Complexos Endossomais de Distribuição Requeridos para Transporte/fisiologia , Herpesvirus Humano 1/fisiologia , Membrana Nuclear/fisiologia , Liberação de Vírus , Animais , Chlorocebus aethiops , Cães , Drosophila melanogaster , Células HEK293 , Células HeLa , Humanos , Células Madin Darby de Rim Canino , Nucleocapsídeo/metabolismo , Ligação Proteica/fisiologia , Coelhos , Células Vero
17.
J Virol ; 92(18)2018 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-29976672

RESUMO

Herpes simplex virus 1 (HSV-1) UL51 is a phosphoprotein that functions in the final envelopment in the cytoplasm and viral cell-cell spread, leading to efficient viral replication in cell cultures. To clarify the mechanism by which UL51 is regulated in HSV-1-infected cells, we focused on the phosphorylation of UL51. Mass spectrometry analysis of purified UL51 identified five phosphorylation sites in UL51. Alanine replacement of one of the identified phosphorylation sites in UL51, serine 184 (Ser-184), but not the other identified phosphorylation sites, significantly reduced viral replication and cell-cell spread in HaCaT cells. This mutation induced membranous invaginations adjacent to the nuclear membrane, the accumulation of primary enveloped virions in the invaginations and perinuclear space, and mislocalized UL34 and UL31 in punctate structures at the nuclear membrane; however, it had no effect on final envelopment in the cytoplasm of HaCaT cells. Of note, the alanine mutation in UL51 Ser-184 significantly reduced the mortality of mice following ocular infection. Phosphomimetic mutation in UL51 Ser-184 partly restored the wild-type phenotype in cell cultures and in mice. Based on these results, we concluded that some UL51 functions are specifically regulated by phosphorylation at Ser-184 and that this regulation is critical for HSV-1 replication in cell cultures and pathogenicity in vivoIMPORTANCE HSV-1 UL51 is conserved in all members of the Herpesviridae family. This viral protein is phosphorylated and functions in viral cell-cell spread and cytoplasmic virion maturation in HSV-1-infected cells. Although the downstream effects of HSV-1 UL51 have been clarified, there is a lack of information on how this viral protein is regulated as well as the significance of the phosphorylation of this protein in HSV-1-infected cells. In this study, we show that the phosphorylation of UL51 at Ser-184 promotes viral replication, cell-cell spread, and nuclear egress in cell cultures and viral pathogenicity in mice. This is the first report to identify the mechanism by which UL51 is regulated as well as the significance of UL51 phosphorylation in HSV-1 infection. Our study may provide insights into the regulatory mechanisms of other herpesviral UL51 homologs.


Assuntos
DNA Helicases/química , DNA Helicases/fisiologia , DNA Primase/química , DNA Primase/fisiologia , Herpesvirus Humano 1/patogenicidade , Proteínas Virais/química , Proteínas Virais/fisiologia , Liberação de Vírus , Replicação Viral , Transporte Ativo do Núcleo Celular , Animais , Linhagem Celular , Chlorocebus aethiops , DNA Helicases/genética , DNA Helicases/isolamento & purificação , DNA Primase/genética , DNA Primase/isolamento & purificação , Olho/virologia , Células HEK293 , Herpes Simples/virologia , Herpesvirus Humano 1/genética , Herpesvirus Humano 1/fisiologia , Humanos , Camundongos , Fosforilação , Proteínas Serina-Treonina Quinases , Células Vero , Proteínas Virais/genética , Proteínas Virais/isolamento & purificação , Vírion/fisiologia , Virulência , Montagem de Vírus
18.
Clin Nephrol Case Stud ; 6: 16-20, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30009125

RESUMO

Thrombocytopenia, anasarca, fever, renal insufficiency, and organomegaly constitute TAFRO syndrome, a variant of Castleman disease. We describe a patient with TAFRO syndrome who underwent renal biopsy. A 79-year-old woman was referred to us with fever and leg edema. She also had thrombocytopenia, pleural effusion, ascites, and acute kidney injury, and was admitted to our hospital. Her response to initial therapy with corticosteroid and cyclosporine was poor. Therefore, she received 4 doses of rituximab per week, which resulted in clinical improvement, including recovery of thrombocytopenia. A kidney biopsy thereafter showed diffuse, global glomerular endothelial injury indicating thrombotic microangiopathy (TMA). These findings suggested that TMA is associated with the thrombocytopenia and renal insufficiency of TAFRO syndrome.

20.
J Virol ; 92(17)2018 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-29899106

RESUMO

UL13 proteins are serine/threonine protein kinases encoded by herpes simplex virus 1 (HSV-1) and HSV-2. Although the downstream effects of the HSV protein kinases, mostly those of HSV-1 UL13, have been reported, there is a lack of information on how these viral protein kinases are regulated in HSV-infected cells. In this study, we used a large-scale phosphoproteomic analysis of HSV-2-infected cells to identify a physiological phosphorylation site in HSV-2 UL13 (i.e., Ser-18) and investigated the significance of phosphorylation of this site in HSV-2-infected cell cultures and mice. Our results were as follows. (i) An alanine substitution at UL13 Ser-18 (S18A) significantly reduced HSV-2 replication and cell-to-cell spread in U2OS cells to a level similar to those of the UL13-null and kinase-dead mutations. (ii) The UL13 S18A mutation significantly impaired phosphorylation of a cellular substrate of this viral protein kinase in HSV-2-infected U2OS cells. (iii) Following vaginal infection of mice, the UL13 S18A mutation significantly reduced mortality, HSV-2 replication in the vagina, and development of vaginal disease to levels similar to those of the UL13-null and the kinase-dead mutations. (iv) A phosphomimetic substitution at UL13 Ser-18 significantly restored the phenotype observed with the UL13 S18A mutation in U2OS cells and mice. Collectively, our results suggested that phosphorylation of UL13 Ser-18 regulated UL13 function in HSV-2-infected cells and that this regulation was critical for the functional activity of HSV-2 UL13 in vitro and in vivo and also for HSV-2 replication and pathogenesis.IMPORTANCE Based on studies on cellular protein kinases, it is obvious that the regulatory mechanisms of protein kinases are as crucial as their functional consequences. Herpesviruses each encode at least one protein kinase, but the mechanism by which these kinases are regulated in infected cells remains to be elucidated, with a few exceptions, although information on their functional effects has been accumulating. In this study, we have shown that phosphorylation of the HSV-2 UL13 protein kinase at Ser-18 regulated its function in infected cells, and this regulation was critical for HSV-2 replication and pathogenesis in vivo UL13 is conserved in all members of the family Herpesviridae, and this is the first report clarifying the regulatory mechanism of a conserved herpesvirus protein kinase that is involved in viral replication and pathogenesis in vivo Our study may provide insight into the regulatory mechanisms of the other conserved herpesvirus protein kinases.


Assuntos
Herpesvirus Humano 2/fisiologia , Proteínas Quinases/metabolismo , Processamento de Proteína Pós-Traducional , Animais , Linhagem Celular , Análise Mutacional de DNA , Modelos Animais de Doenças , Herpes Genital/patologia , Herpes Genital/virologia , Herpesvirus Humano 2/genética , Herpesvirus Humano 2/patogenicidade , Humanos , Camundongos , Fosforilação , Proteínas Quinases/genética , Internalização do Vírus , Liberação de Vírus , Replicação Viral
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA