Nuclear translocation of mutagenized forms of human cytomegalovirus glycoprotein B (gpUL55).

Bogner, Elke; Anheier, Bärbel; Offner, Fritz; Smuda, Christoph; Reschke, M.; Eickmann, Markus; Radsak, K.

doi:10.1099/0022-1317-78-7-1647

Cited by 8 publications

(5 citation statements)

References 18 publications

(24 reference statements)

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“…2A, a) characteristic staining was obtained of the RER and the nuclear envelope (Fig. 2B, a), which corresponded to that in infected or stably transfected cells described previously (24). Immunoprecipitations confirmed the presence of HCMV gB in solubilized nuclear fractions representing INM as well as postnuclear fractions representing solubilized cytoplasmic membranes including RER and plasma membranes (Fig.…”

Section: The Carboxyl-terminal Domain Of the Cytoplasmic Tail Of Hcmvsupporting

confidence: 81%

“…previous experiments have shown that translocation of recombinant HCMV gB to the INM was dependent on the 51 carboxyl-terminal amino acids (24). In order to address the question of whether the putative signal within this domain was transferable, chimeras of defined segments of HCMV gB and the plasma membrane protein CD8 as a reporter were constructed ( Fig.…”

Section: The Carboxyl-terminal Domain Of the Cytoplasmic Tail Of Hcmvmentioning

confidence: 99%

“…Our previous experiments have shown that in the case of HCMV gB, the nucleoplasmic COOH-terminal 51 amino acids are essential for targeting to the INM (24).…”

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confidence: 99%

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Identification of a Novel Signal Sequence That Targets Transmembrane Proteins to the Nuclear Envelope Inner Membrane

Meyer

Radsak

2000

Journal of Biological Chemistry

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Herpesvirus maturation requires translocation of glycoprotein B homologue from the endoplasmic reticulum to the inner nuclear membrane. Glycoprotein B of human cytomegalovirus was used in this context as a model protein. To identify a specific signal sequence within human cytomegalovirus glycoprotein B acting in a modular fashion, coding sequences were recombined with reporter proteins. Immunofluorescence and cell fractionation demonstrated that a short sequence element within the cytoplasmic tail of human cytomegalovirus glycoprotein B was sufficient to translocate the membrane protein CD8 to the inner nuclear membrane. This carboxyl-terminal sequence had no detectable nuclear localization signal activity for soluble ␤-Galactosidase and could not be substituted by the nuclear localization signal of SV40 T antigen. For glycoprotein B of herpes simplex virus, a carboxyl-terminal element with comparable properties was found. Further experiments showed that the amino acid sequence DRLRHR of human cytomegalovirus glycoprotein B (amino acids 885-890) was sufficient for nuclear envelope translocation. Single residue mutations revealed that the arginine residues in positions 4 and 6 of the DRLRHR sequence were essential for its function. These results support the view that transmembrane protein transport to the inner nuclear membrane is controlled by a mechanism different from that of soluble proteins.The nuclear envelope consists of three morphologically distinct membrane domains (1, 2): (i) the outer nuclear membrane, which is contiguous with the membranes of the rough endoplasmic reticulum (RER), 1 (ii) the pore membranes, which connect outer and inner nuclear membranes and are associated with the nuclear pore complexes, and (iii) the inner nuclear membrane (INM), which is adjacent to the nuclear lamina, a mesh-work of intermediate filament proteins termed lamins (3). Inner and outer nuclear membranes enclose the perinuclear space that continues into that of the RER. While the outer nuclear membrane shares the membrane proteins of the RER, the INM contains a distinct set of integral proteins (4), including the lamin B receptor (LBR) (5) and the lamina-associated polypeptides 1 and 2 (LAP1 and LAP2) (6, 7).In cells that are virus-infected, the various cellular membranes are decorated with specific viral proteins in addition. In the case of herpesviruses that undergo a characteristic nuclear phase of maturation, viral transmembrane glycoproteins are also translocated from the site of their synthesis into the INM, e.g. the glycoprotein B (gB) homologues of Epstein-Barr virus (8, 9), herpes simplex virus type 1 (HSV-1) (10), and human cytomegalovirus (HCMV) (11). The specific localization in the INM not only suggests a role of these proteins in herpesvirus envelopment at the inner nuclear membrane (12) but also implies that specific mechanisms mediate their transport from the RER to the INM compartment. It is currently not precisely known how this apparently specific targeting of cellular and viral transmembrane protein...

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Section: The Carboxyl-terminal Domain Of the Cytoplasmic Tail Of Hcmvsupporting

confidence: 81%

Section: The Carboxyl-terminal Domain Of the Cytoplasmic Tail Of Hcmvmentioning

confidence: 99%

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Identification of a Novel Signal Sequence That Targets Transmembrane Proteins to the Nuclear Envelope Inner Membrane

Meyer

Radsak

2000

Journal of Biological Chemistry

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“…gB is a type I glycoprotein containing a signal sequence, an extracellular or luminal domain, a transmembrane (TM) domain, and a 135-amino-acid cytoplasmic tail (5,20,38). The cytoplasmic tail contains a consensus casein kinase II (CKII) site, which is phosphorylated both in vitro and in vivo (1,37,55).…”

mentioning

confidence: 99%

Steady-State Plasma Membrane Expression of Human Cytomegalovirus gB Is Determined by the Phosphorylation State of Ser ₉₀₀

Fish

Söderberg‐Nauclér

Nelson

1998

J Virol

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Human cytomegalovirus (HCMV) infection of an astrocytoma cell line (U373) or human fibroblast (HF) cells results in a differential cell distribution of the major envelope glycoprotein gB (UL55). This 906-amino-acid type I glycoprotein contains an extracellular domain with a signal sequence, a transmembrane domain, and a 135-amino-acid cytoplasmic tail with a consensus casein kinase II (CKII) site located at Ser900. Since phosphorylation of proteins in the secretory pathway is an important determinant of intracellular trafficking, the state of gB phosphorylation in U373 and HF cells was examined. Analysis of cells expressing wild-type gB and gB with site-specific mutations indicated that the glycoprotein was equally phosphorylated at a single site, Ser900, in both U373 and HF cells. To assess the effect of charge on gB surface expression in U373 cells, Ser900 was replaced with an aspartate (Asp) or alanine (Ala) residue to mimic the phosphorylated and nonphosphorylated states, respectively. Expression of the Asp but not the Ala gB mutation resulted in an increase in the steady-state expression of gB at the plasma membrane (PM) in U373 cells. In addition, treatment of U373 cells with the phosphatase inhibitor tautomycin resulted in the accumulation of gB at the PM. Interestingly, the addition of a charge at Ser900 trapped gB in a low-level cycling pathway at the PM, preventing trafficking of the protein to thetrans-Golgi network or other intracellular compartments. Therefore, these results suggest that a tautomycin-sensitive phosphatase regulates cell-specific PM retrieval of gB to intracellular compartments.

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“…In addition to the nuclear localization signal, the cytoplasmic domain also contains several internalization signals for retrieval from the infected cell surface for virus envelopment [9,14,20,21]. Among the various internalization signals there are the tyrosine motifs Y 845 QML and Y 894 RHL, the dileucine motifs LL 849 and LL 884 , and an acid sequence containing serine 900 , which may be phosphorylated by casein kinase II and thus modulate the retrieval of gB from the plasma membrane [2,9,21] (Fig. 1).…”

Section: Introductionmentioning

confidence: 99%

Multimerization potential of the cytoplasmic domain of the human cytomegalovirus glycoprotein B

Scheffczik¹,

Kraus²,

Kiermayer³

et al. 2001

FEBS Letters

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In the present study the coding sequence of the cytoplasmic tail of the human cytomegalovirus glycoprotein B (gB) was expressed. The secondary structure of the purified recombinant protein was analyzed by circular dichroism, and the quaternary structure was investigated by gel permeation chromatography, and electron microscopy. Our data indicate that the cytoplasmic gB domain contains K K-helix structures and assembles into tetramers, suggesting that the authentic gB may represent a homotetramer. ß

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Nuclear translocation of mutagenized forms of human cytomegalovirus glycoprotein B (gpUL55).

Cited by 8 publications

References 18 publications

Identification of a Novel Signal Sequence That Targets Transmembrane Proteins to the Nuclear Envelope Inner Membrane

Identification of a Novel Signal Sequence That Targets Transmembrane Proteins to the Nuclear Envelope Inner Membrane

Steady-State Plasma Membrane Expression of Human Cytomegalovirus gB Is Determined by the Phosphorylation State of Ser ₉₀₀

Multimerization potential of the cytoplasmic domain of the human cytomegalovirus glycoprotein B

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Nuclear translocation of mutagenized forms of human cytomegalovirus glycoprotein B (gpUL55).

Cited by 8 publications

References 18 publications

Identification of a Novel Signal Sequence That Targets Transmembrane Proteins to the Nuclear Envelope Inner Membrane

Identification of a Novel Signal Sequence That Targets Transmembrane Proteins to the Nuclear Envelope Inner Membrane

Steady-State Plasma Membrane Expression of Human Cytomegalovirus gB Is Determined by the Phosphorylation State of Ser 900

Multimerization potential of the cytoplasmic domain of the human cytomegalovirus glycoprotein B

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Steady-State Plasma Membrane Expression of Human Cytomegalovirus gB Is Determined by the Phosphorylation State of Ser ₉₀₀