Posttranslational modifications of Sindbis virus glycoproteins: electrophoretic analysis of pulse-chase-labeled infected cells

Bonatti, Stefano; Cancedda, Fiorella Descalzi

doi:10.1128/jvi.42.1.64-70.1982

Cited by 29 publications

(8 citation statements)

References 29 publications

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“…The electrophoretic mobility of E2 was, however, significantly increased upon digestion with endo F (lane 3). These data are consistent with published reports showing that PE2 contains highmannose oligosaccharides (2,9), while E2 contains complex oligosaccharides in combination with oligosaccharides which are not accessible to enzymatic alteration and therefore remain in the high-mannose form (11). When the PE2 glycoprotein produced in RPE.40 cells was treated with endo H, its mobility was increased, as was that of endo H-treated PE2 from CHO-Ki cells.…”

Section: 4supporting

confidence: 93%

A mutant CHO-K1 strain with resistance to Pseudomonas exotoxin A and alphaviruses fails to cleave Sindbis virus glycoprotein PE2

Watson

Moehring

1991

J Virol

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RPE.40, a mutant CHO-Kl strain selected for resistance to Pseudomonas exotoxin A, is defective in the production of infectious alphaviruses, although viruses are taken in and processed normally (J. M. Moehring and T. J. Moehring, Infect. Immun. 41:998-1009, 1983). To determine the cause of this defect, the synthesis of Sindbis virus proteins was examined. RPE.40 cells produced and glycosylated structural glycoprotein precursors PE2 and immature El normally. Mature El was formed, but PE2 was not cleaved to E2 and E3. PE2 instead was modified to a higher-molecular-weight form (PE2') in which the high-mannose oligosaccharides were processed to the complex form without proteolytic cleavage. The data suggest that the cleavage which produces E2 occurs within the trans-Golgi or in post-Golgi elements and is closely associated with the addition of sialic acid residues to the asparagine-linked oligosaccharides. RPE.40 cells make and release noninfectious Sindbis virions that contain PE2' and no detectable E2. These virions can be converted to an infectious form by treatment with trypsin. A defect in an intracellular endopeptidase activity in RPE.40 cells is postulated. Comparison of two Sindbis virus strains showed that the requirement for E2 in the virion to ensure infectivity is strain specific.

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Section: 4supporting

confidence: 93%

A mutant CHO-K1 strain with resistance to Pseudomonas exotoxin A and alphaviruses fails to cleave Sindbis virus glycoprotein PE2

Watson

Moehring

1991

J Virol

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“…16). E l and pE 2 are glycosylated co-translationally in the endoplasmic reticulum, and while moving from the endoplasmic reticulum, through the Golgi system, to the plasma membrane, pE2 and EL form a complex, their oligosaccharides are processed and pE2 is cleaved, presumably in the Golgi membranes, by a trypsin-like enzyme to E2 and E3 (Keegstra and Burke 1975;Bell and Strauss, 1981;Schmidt and Schlessinger, 1982;Griffiths et al, 1983;Tabas and Kornfeld, 1978;Bonatti and Cancedda, 1982;Ziemiecki et al, 1980). This proteolytic cleavage of pE 2 is essential for virus budding .…”

Section: Sindbis Virusmentioning

confidence: 99%

Inhibitors of protein glycosylation and glycoprotein processing in viral systems

Datema

Olofsson²,

Romero

1987

Pharmacology & Therapeutics

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“…SDS gel electrophoresis was performed as described by Laemmli ( 22) and modified by Bonatti and Descalzi-Cancedda (5). The concentration of acrylamide was 9%.…”

Section: Pagementioning

confidence: 99%

Type X collagen synthesis during in vitro development of chick embryo tibial chondrocytes.

Castagnola¹,

Moro²,

Descalzi-Cancedda³

et al. 1986

The Journal of Cell Biology

133

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Abstract. In the developing chick embryo tibia type X collagen is synthesized by chondrocytes from regions of hypertrophy and not by chondrocytes from other regions (Capasso, O., G. Tajana, and R. Cancedda, 1984, Mol. Cell. Biol. 4:1163-1168 Schmid, T. M., and T. F. Linsenmayer, 1985, Dev. Biol. 107:375-381). To investigate further the relationship between differentiation of endochondral chondrocytes and type X collagen synthesis we have developed a novel culture system for chondrocytes from 29-31-stage chick embryo tibiae. At the beginning of the culture these chondrocytes are small and synthesize type II and not type X collagen, but when grown on agarose-coated dishes they further differentiate into hypertrophic chondrocytes that synthesize type X collagen. The synthesis of type X collagen has been monitored in cultured cells by analysis of labeled collagens and in vitro translation of mRNAs. When the freshly dissociated chondrocytes are plated in anchorage-permissive dishes, most of the cells attach and dedifferentiate, as revealed by their fibroblastic morphology. Dedifferentiated chondrocytes, after several passages, can still reexpress the differentiated phenotype and continue their development to hypertrophic, type X collagen-synthesizing chondrocytes. Hypertrophic chondrocytes, when plated in anchorage permissive dishes, attach, maintaining the differentiated phenotype, and continue the synthesis of type X collagen.T HE development and growth of long bones mainly occurs by endochondrai bone formation. During this process, chondrocytes in the bone growth region pass sequentially through a proliferative, a hypertrophic, and a degenerative stage (44). Differentiating chondrocytes synthesize collagens and other extracellular matrix macromolecules which are deposited in the cartilage. For several years type II collagen has been recognized as the only cartilage-specific collagen (29). More recently the existence of other minor collagens has been demonstrated, including 1 c~, 2a, 3a chains (6), and type IX (33, 34, 45) and X collagens. In particular, type X collagen, first detected in cultures of chick embryo tibial chondrocytes (8,9,37,38) and in cultures of chick embryo sternal chondrocytes grown within collagen gels (16, 17) and subsequently in other species (35), becomes a major collagen in the regions where cartilage is removed and replaced by bone tissue. Its synthesis by hypertrophic chondrocytes and its association with endochondral development have been shown by metabolic labeling of proteins made by the cells (10,20) and by immunofluorescent staining of cartilage with specific monoclonal antibodies (40). From the same regions of the tibiae the type X collagen deposited in the cartilage in vivo has been purified in milligram quantities (9,32). After performing cultures of chondrocytes from sterna of chick embryos at different stages of development, Gibson and Flint (14) reported synthesis of type X collagen by chondrocytes derived from the presumptive calcification region but not by chondrocytes der...

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Posttranslational modifications of Sindbis virus glycoproteins: electrophoretic analysis of pulse-chase-labeled infected cells

Cited by 29 publications

References 29 publications

A mutant CHO-K1 strain with resistance to Pseudomonas exotoxin A and alphaviruses fails to cleave Sindbis virus glycoprotein PE2

A mutant CHO-K1 strain with resistance to Pseudomonas exotoxin A and alphaviruses fails to cleave Sindbis virus glycoprotein PE2

Inhibitors of protein glycosylation and glycoprotein processing in viral systems

Type X collagen synthesis during in vitro development of chick embryo tibial chondrocytes.

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