Identification of a novel functional nuclear localization signal in the protein encoded by open reading frame 47 of Bombyx mori nucleopolyhedrovirus

2010 ◽  
Vol 155 (12) ◽  
pp. 1943-1950 ◽  
Author(s):  
Zhong-Jian Guo ◽  
Dian-Xuan Wang ◽  
Qin Yao ◽  
Ke-Ping Chen ◽  
Chuan-Xi Zhang
Keyword(s):  
Bombyx Mori ◽  
Nuclear Localization ◽  
Nuclear Localization Signal ◽  
Open Reading Frame ◽  
Bombyx Mori Nucleopolyhedrovirus ◽  
Reading Frame ◽  
Localization Signal ◽  
Functional Nuclear Localization Signal

scholarly journals Regions of the Varicella-Zoster Virus Open Reading Frame 63 Latency-Associated Protein Important for Replication In Vitro Are Also Critical for Efficient Establishment of Latency

2005 ◽  
Vol 79 (8) ◽  
pp. 5069-5077 ◽  
Author(s):  
Jeffrey I. Cohen ◽  
Tammy Krogmann ◽  
Sebastien Bontems ◽  
Catherine Sadzot-Delvaux ◽  
Lesley Pesnicak
Keyword(s):  
Amino Acids ◽  
Varicella Zoster Virus ◽  
Nuclear Localization ◽  
Nuclear Localization Signal ◽  
Open Reading Frame ◽  
Reading Frame ◽  
Varicella Zoster ◽  
Localization Signal ◽  
Carboxy Terminal

ABSTRACT Varicella-zoster virus (VZV) open reading frame 63 (ORF63) is one of the most abundant transcripts expressed during VZV latency in humans, and ORF63 protein has been detected in human ganglia by several laboratories. Deletion of over 90% of the ORF63 gene showed that the protein is required for efficient establishment of latency in rodents. We have constructed viruses with a series of mutations in ORF63. While prior experiments showed that transfection of cells with a plasmid expressing ORF63 but lacking the putative nuclear localization signal of the protein resulted in increased expression of the protein in the cytoplasm, we found that ORF63 protein remained in the nucleus in cells infected with a VZV ORF63 nuclear localization signal deletion mutant. This mutant was not impaired for growth in cell culture or for latency in rodents. Replacement of five serine or threonine phosphorylation sites in ORF63 with alanines resulted in a virus that was impaired for replication in vitro and for latency. A series of ORF63 carboxy-terminal mutants showed that the last 70 amino acids do not affect replication in vitro or latency in rodents; however, the last 108 amino acids are important for replication and latency. Thus, regions of ORF63 that are important for replication in vitro are also required for efficient establishment of latency.

scholarly journals Dissection of a Novel Nuclear Localization Signal in Open Reading Frame 29 of Varicella-Zoster Virus

2005 ◽  
Vol 79 (20) ◽  
pp. 13070-13081 ◽  
Author(s):  
Christina L. Stallings ◽  
Saul Silverstein
Keyword(s):  
Dorsal Root Ganglia ◽  
Nuclear Localization ◽  
Nuclear Import ◽  
Nuclear Localization Signal ◽  
Dorsal Root ◽  
Open Reading Frame ◽  
Nuclear Targeting ◽  
Reading Frame ◽  
Varicella Zoster ◽  
Localization Signal

ABSTRACT Open reading frame 29 (ORF29) of varicella-zoster virus (VZV) encodes a 120-kDa single-stranded DNA binding protein (ORF29p) that is not packaged in the virion and is expressed during latency. During lytic infection, ORF29p is localized primarily to infected cell nuclei. In contrast, ORF29p is found exclusively in the cytoplasm in neurons of the dorsal root ganglia obtained at autopsy from seropositive latently infected patients. ORF29p accumulates in the nuclei of neurons in dorsal root ganglia obtained at autopsy from patients with active zoster. The localization of this protein is, therefore, tightly correlated with the proposed VZV lytic/latent switch. In this report, we have investigated the nuclear import mechanism of ORF29p. We identified a novel nuclear targeting domain bounded by amino acids 9 to 154 of ORF29p that functions independent of other VZV-encoded factors. In vitro import assays in digitonin-permeabilized HeLa cells reveal that ORF29p is transported into the nucleus by a Ran-, karyopherin α- and β-dependent mechanism. These data are further supported by the demonstration that a glutathione S-transferase-karyopherin α fusion interacts with ORF29p, but not with a protein containing a point mutation in its nuclear localization signal (NLS). Therefore, the region of ORF29p responsible for its nuclear targeting is also involved in the association with karyopherin α. As a result of this interaction, this noncanonical NLS appears to hijack the classical cellular nuclear import machinery. Elucidation of the mechanisms governing ORF29p nuclear targeting could shed light on the VZV reactivation process.

Characterization of the open reading frame 7a from Bombyx mori nucleopolyhedrovirus

2012 ◽  
Vol 40 (2) ◽  
pp. 865-873
Author(s):  
Yong Liu ◽  
Feng Yu ◽  
Huiling Wu ◽  
Qing Cao ◽  
Yu Wu ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Open Reading Frame ◽  
Bombyx Mori Nucleopolyhedrovirus ◽  
Reading Frame

Open reading frame 60 of the Bombyx mori nucleopolyhedrovirus plays a role in budded virus production

2010 ◽  
Vol 151 (2) ◽  
pp. 185-191 ◽  
Author(s):  
Zhong-Jian Guo ◽  
Li-Hua Qiu ◽  
Shi-Heng An ◽  
Qin Yao ◽  
Enoch Y. Park ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Virus Production ◽  
Open Reading Frame ◽  
Bombyx Mori Nucleopolyhedrovirus ◽  
Reading Frame ◽  
Budded Virus

scholarly journals SUMOylation of the Polyglutamine Repeat Protein, Ataxin-1, Is Dependent on a Functional Nuclear Localization Signal

2005 ◽  
Vol 280 (23) ◽  
pp. 21942-21948 ◽  
Author(s):  
Brigit E. Riley ◽  
Huda Y. Zoghbi ◽  
Harry T. Orr
Keyword(s):  
Nuclear Localization ◽  
Nuclear Localization Signal ◽  
High Mobility ◽  
Wild Type ◽  
Repeat Protein ◽  
Polyglutamine Repeat ◽  
Lysine Residues ◽  
Localization Signal ◽  
Functional Nuclear Localization Signal ◽  
Polyglutamine Tract

SUMO (small ubiquitin-like modifier) is a member of the ubiquitin family of proteins. SUMO targets include proteins involved in numerous roles including nuclear transport and transcriptional regulation. The previous finding that mutant ataxin-1[82Q] disrupted promyelocytic leukemia (PML) oncogenic domains prompted us to determine whether ataxin-1 disrupts another component of PML oncogenic domains, Sp100 (100-kDa Speckled protein). Similar to the PML protein, mutant ataxin-1[82Q] redistributed Sp100 to mutant ataxin-1[82Q] nuclear inclusions. Based on the ability of PML and Sp100 to be covalently modified by SUMO, we investigated the ability of ataxin-1 to be SUMOylated. SUMO-1 was found to covalently modify the polyglutamine repeat protein ataxin-1. There was a decrease in ataxin-1 SUMOylation in the presence of the expanded polyglutamine tract, ataxin-1[82Q]. The phospho-mutant, ataxin-1[82Q]-S776A, restored SUMO levels to those of wild-type ataxin-1[30Q]. SUMOylation of ataxin-1 was dependent on a functional nuclear localization signal. Ataxin-1 SUMOylation was mapped to at least five lysine residues. Lys16, Lys194 preceding the polyglutamine tract, Lys610/Lys697 in the C-terminal ataxin high mobility group domain, and Lys746 all contribute to ataxin-1 SUMOylation.

scholarly journals Properties of expression of protein capside porcine circovirus type 2 in bacterial cells

2021 ◽  
pp. 48-57
Author(s):  
Anastasia D. Titova ◽  
Kirill V. Kudzin ◽  
Vladimir A. Prokulevich
Keyword(s):  
Nuclear Localization ◽  
Nuclear Localization Signal ◽  
Porcine Circovirus Type ◽  
Porcine Circovirus Type 2 ◽  
Porcine Circovirus ◽  
Bacterial Cells ◽  
Reading Frame ◽  
E Coli ◽  
Localization Signal

To improve expression of the porcine circovirus type 2 (PCV2) capsid protein in E. coli cells, the corresponding gene was optimized and two variants of the open reading frame were constructed, which encoded the full-sized and shortened capsid proteins as part of the expression vector. Rare codons were replaced, and in the case of a shortened version of the gene, the region corresponding to the N-terminal domain of the protein was deleted. A comparison was made of the expression level of the studied proteins. It was established that the highest level of expression in bacterial cells is achieved by simultaneously optimizing the codons and removing the initial (N-terminal) 108 base pair (bp) portion of the gene, which contains the nuclear localization signal.

scholarly journals Human Cytomegalovirus UL84 Localizes to the Cell Nucleus via a Nuclear Localization Signal and Is a Component of Viral Replication Compartments

2002 ◽  
Vol 76 (17) ◽  
pp. 8931-8938 ◽  
Author(s):  
Yiyang Xu ◽  
Kelly S. Colletti ◽  
Gregory S. Pari
Keyword(s):  
Amino Acids ◽  
Viral Replication ◽  
Human Cytomegalovirus ◽  
Nuclear Localization ◽  
Nuclear Localization Signal ◽  
Human Fibroblasts ◽  
Reading Frame ◽  
Early Protein ◽  
Localization Signal ◽  
Infected Cells

ABSTRACT The UL84 open reading frame encodes a protein that is required for origin-dependent DNA replication and interacts with the immediate-early protein IE2 in lytically infected cells. Transfection of UL84 expression constructs showed that UL84 localized to the nucleus of transfected cells in the absence of any other viral proteins and displayed a punctate speckled fluorescent staining pattern. Cotransfection of all the human cytomegalovirus replication proteins and oriLyt, along with pUL84-EGFP, showed that UL84 colocalized with UL44 (polymerase accessory protein) in replication compartments. Experiments using infected human fibroblasts demonstrated that UL84 also colocalized with UL44 and IE2 in viral replication compartments in infected cells. A nuclear localization signal was identified using plasmid constructs expressing truncation mutants of the UL84 protein in transient transfection assays. Transfection assays showed that UL84 failed to localize to the nucleus when 200 amino acids of the N terminus were deleted. Inspection of the UL84 amino acid sequence revealed a consensus putative nuclear localization signal between amino acids 160 and 171 (PEKKKEKQEKK) of the UL84 protein.

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