Multiplicity dependent expression of the predominant phosphoprotein pp65 of human cytomegalovirus

Klages, Sabine; Rüger, Barbara; Jahn, Gerhard

doi:10.1016/0168-1702(89)90061-0

Cited by 28 publications

(22 citation statements)

References 20 publications

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“…Our results with native and recombinant pp65 confirm other studies in which the humoral immune response against pp65 was found to be highly variable and sometimes absent (Landini et al, 1985;Jahn et al, 1987;Klages et al, 1989;Plachter et al, 1990;Pande et al, 1991). Using the insect-expressed antigens, we were able to prove that the epitopes recognized by MAbs 4D11, 7D2, 8El0 and 7B4 mapped in the region of pp65, comprising amino acids 1 to 442, and that the phosphoprotein GP66 (Kim et al, 1983) and the 65K phosphoprotein are the same.…”

supporting

confidence: 82%

“…This phosphoprotein has also been designated the lower matrix protein (Gibson, 1983), gp64 (Gibson, 1983;Clark et al, 1984;Forman et al, 1985;Pande et al, 1984) and ICP27 (Geballe et al, 1986). Numerous reports have described pp65 as one of the HCMV proteins most frequently recognized on Western blots by human sera from individuals seropositive for the virus (Landini et al, 1985;Jahn et al, 1987;Klages et al, 1989;Landini & LaPlaca, 1991).…”

mentioning

confidence: 99%

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Expression of the human cytomegalovirus 65K tegument phosphoprotein in insect cells by baculovirus vectors

Fauci¹,

Sapienza²,

Chen³

et al. 1994

Journal of General Virology

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The gene encoding the 65K tegument phosphoprotein (pp65) of human cytomegalovirus (HCMV) was cloned into pAc373 to construct a recombinant baculovirus (Acpp65-3) expressing pp65 in insect Sf9 cells. A baculovirus that carried a fragment of the gene, corresponding to the first 442 amino acids ofpp65, was also developed, using vector pVL941 (Acpp65-2). Recombinant proteins migrating in SDSpolyacrylamide gels with an M r of either 65K (Acpp65-3) or 56K (Acpp65-2) were detected in cytoplasmic and nuclear extracts of infected Sf9 cells. The 56K and 65K proteins were recognized in immunoblots by monoclonal antibodies (MAbs) 28-77 and 28-19, which are specific for pp65. The insect cell-expressed antigens were also analysed on Western blots using MAbs 4D11, 7D2, 8E3, 7B4 and 8El0, which recognize the HCMV antigen GP66 in immunoblots. The truncated pp65 antigen of Acpp65-2 was reactive with MAbs 4Dll, 7D2, 8El0 and 7B4. The protein expressed by Acpp65-3 reacted only with MAb 4D11. The data proved that the epitopes recognized by MAbs 4Dll, 7D2, 8E3 and 7B4 mapped in the region ofpp65, comprising amino acids 1 to 442, and also that GP66 and pp65 represent the same HCMV antigen. Immunoblot analysis of human sera from individuals seropositive for HCMV showed that the recombinant pp65 products were as antigenic as the native 65K phosphoprotein produced in HCMVinfected human embryonic fibroblasts.

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supporting

confidence: 82%

mentioning

confidence: 99%

Expression of the human cytomegalovirus 65K tegument phosphoprotein in insect cells by baculovirus vectors

Fauci¹,

Sapienza²,

Chen³

et al. 1994

Journal of General Virology

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“…Recent clinical isolates of HCMV display considerable quantitative variations in the expression of ppUL83 (SiqueiraLinhares et al, 1981;Klages et al, 1989). Klages et al (1989) found that a reduced production of ppUL83 correlates with reduced amounts of its transcript in wildtype strains compared to the high level of expression found in the laboratory strain AD 169. Other studies do not seem to support this finding completely (FauconBiguet et al, 1986;Grefte et al, 1993).…”

Section: (Iii) Pp Ul82 and (Iv) Pp Ul83mentioning

confidence: 99%

“…A MAb (9220) and immunoelectron microscopy were used to localize ppUL83 to the matrix of the dense bodies (Landini et al, 1987). Recent clinical isolates of HCMV display considerable quantitative variations in the expression of ppUL83 (SiqueiraLinhares et al, 1981;Klages et al, 1989). Klages et al (1989) found that a reduced production of ppUL83 correlates with reduced amounts of its transcript in wildtype strains compared to the high level of expression found in the laboratory strain AD 169.…”

Section: (Iii) Pp Ul82 and (Iv) Pp Ul83mentioning

confidence: 99%

Human cytomegalovirus structural proteins

Spaete¹,

Gehrz²,

Landini³

1994

Journal of General Virology

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“…Another abundantly produced tegument protein of HCMV is pp65, which is also called the lower matrix protein and is inappropriately over-expressed by cultured fibroblasts with increasing passage (Geballe et al, 1986;Klages et al, 1989). Dense bodies, non-infectious particles that accumulate in the cytoplasm of infected cells in culture, contain an abundance of the tegument protein pp65, accounting for approximately 95% of their protein mass (Fiala et al, 1976;Irmiere & Gibson, 1983).…”

Section: Introductionmentioning

confidence: 99%

Nuclear localization of the human cytomegalovirus tegument protein pp150 (ppUL32)

Hensel

Meyer²,

Gärtner³

et al. 1995

Journal of General Virology

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The human cytomegalovirus (HCMV) basic phosphoprotein pp 150, encoded by the UL32 gene, together with the two other major phosphoproteins, pp65 (ppUL83) and pp71 (ppUL82) and several minor structural proteins, form the tegument around the viral nucleocapsid. Experiments were undertaken to locate the area of assembly of tegument proteins ppl50 and pp65 and nucleocapsids in fibroblasts, in order to assess the functional role of these two structural proteins in HCMV morphogenesis. Whereas ppl50 expression starts during the cytoplasmic maturation of HCMV, pp65 is expressed in the early and late phases of HCMV gene transcription. Western blot analysis of isolated cell fractions showed that ppl50 is initially (48 h postinfection) localized in the nucleus, associated either with the nuclear membrane or with viral assembly regions, and later (72 h post-infection) in the cytoplasm. By indirect immunofluorescence, ppl50 and pp65 could be detected in nuclear subcompartments and were strongly associated with the nuclear membrane. Using immunogold analysis by electron microscopy, pp65 was exclusively detected within the matrix of cytoplasmic and extracellular dense bodies and of dense body-like structures in the nucleoplasm. These were localized in close contact with hypertrophic nucleoli, in the proximity of developing nucleocapsids and in special patches at the inner nuclear membrane L Positive immunostaining of ppl50 was observed at the surface of developing nucleocapsids concentrated within viral assembly regions in the nucleoplasm. Additionally, the tegument of cytoplasmic and extracellular virions was stained, whereas dense bodies or nuclear dense body-like structures did not react. Thus, the acquisition of the tegument protein pp 150 seems to start in special nuclear subcompartments of the HCMV-infected fibroblasts.

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Multiplicity dependent expression of the predominant phosphoprotein pp65 of human cytomegalovirus

Cited by 28 publications

References 20 publications

Expression of the human cytomegalovirus 65K tegument phosphoprotein in insect cells by baculovirus vectors

Expression of the human cytomegalovirus 65K tegument phosphoprotein in insect cells by baculovirus vectors

Human cytomegalovirus structural proteins

Nuclear localization of the human cytomegalovirus tegument protein pp150 (ppUL32)

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