Electron microscopy of the nucleocapsid from disrupted Moloney murine leukemia virus and of associated type VI collagen-like filaments

Pager, Jeanne; Coulaud, Dominique; Delain, Etienne

doi:10.1128/jvi.68.1.223-232.1994

Cited by 12 publications

(7 citation statements)

References 54 publications

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“…Treatment with 0.01% Nonidet P-40 after assembly disrupted the cylindrical particles, but it did not seem to dissociate the protein from the RNA. Instead, what appeared to be large tangles of RNA covered with protein, with many small loops, were produced (data not shown), which resembled the structures observed after the disruption of Moloney MLV by freezing-thawing (45). For RSV as well as other retroviruses it is known that immature cores are stable in the presence of nonionic detergents after the removal of the lipid membrane, while, at least for most retroviruses, mature cores fall apart under the same conditions (58).…”

Section: Resultsmentioning

confidence: 59%

Self-assembly in vitro of purified CA-NC proteins from Rous sarcoma virus and human immunodeficiency virus type 1

Campbell

Vogt

1995

J Virol

333

150

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The internal structural proteins of retroviruses are proteolytically processed from the Gag polyprotein, which alone is able to assemble into virus-like particles when expressed in cells. All Gag proteins contain domains corresponding to the three structural proteins MA, CA, and NC. We have expressed the CA and NC domains together as a unit in Escherichia coli, both for Rous sarcoma virus (RSV) and for human immunodeficiency virus type 1 (HIV-1). We also expressed a similar HIV-1 protein carrying the C-terminal p6 domain. RSV CA-NC, HIV-1 CA-NC, and HIV-1 CA-NC-p6 were purified in native form by classic methods. After adjustment of the pH and salt concentration, each of these proteins was found to assemble at a low level of efficiency into structures that resembled circular sheets and roughly spherical particles. The presence of RNA dramatically increased the efficiency of assembly, and in this case all three proteins formed hollow, cylindrical particles whose lengths were determined by the size of the RNA. The optimal pH at which assembly occurred was 5.5 for the RSV protein and 8.0 for the HIV-1 proteins. The treatment of the RSV CA-NC cylindrical particles with nonionic detergent, with ribonuclease, or with viral protease caused disassembly. These results suggest that RNA plays an important structural role in the virion and that it may initiate and organize the assembly process. The in vitro system described should facilitate the dissection of assembly pathways in retroviruses.

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Section: Resultsmentioning

confidence: 59%

Self-assembly in vitro of purified CA-NC proteins from Rous sarcoma virus and human immunodeficiency virus type 1

Campbell

Vogt

1995

J Virol

333

150

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“…The production of type VI collagen is obviously modified in skin pathologies. A decrease in synthesis, for instance, has been reported in cutis laxa (Crawford et al, 1985), and an increased synthesis is suggested for fibroblasts infected with viruses (Pager et al, 1994). Finally, a different regulatory mechanism for the expression of type VI collagen, compared with fibrillar collagens, has been documented (Hatamochi et al, 1989).…”

Section: Type VI Collagen: a Tying Device For Bundlesmentioning

confidence: 99%

Distribution of minor collagens during skin development

Garrone

Lethias

Guellec

1997

Microsc. Res. Tech.

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The skin is a tissue containing a large number of collagen types. Several collagens are restricted at the dermo‐epidermal junction, contrarily to others present throughout the dermis. However, the distribution of the dermal collagen varies during embryonic development. In this contribution, we have been interested in the collagen types associated with the major collagenous components of the dermis, which are the collagen types I and III. Type V collagen, which is mixed with collagen types I and III to form heterotypic fibrils, has been studied during mouse embryo development. Transcripts of the α1(V) gene have been localized by in situ hybridization, on flattened cells of the stratum germinativum first, and then only on dermal cells. The expression of the gene decreases at birth, while the expression of the α1(I) gene remains constant, with, however, a ring of high intensity around hair follicles. Other collagen types (VI, and the fibril‐associated collagens XII and XIV) have been studied during calf embryonic development by immunofluorescence and ultrastructural immunogold detection. Type VI collagen appears homogeneously distributed throughout the dermis. Type XII collagen is first widely distributed and becomes restricted in the upper, papillary dermis after 6 months of gestation. Type XIV collagen, on the contrary, is first located as a delicate framework around hair follicles (at 19 weeks of gestation), and progressively invades the whole dermis where it appears abundant just before birth. The different functions of all these collagens are discussed in terms of dermis architecture, mechanical properties and physiology. Microsc. Res. Tech. 38:407–412, 1997. © 1997 Wiley‐Liss, Inc.

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“…The retroviral NC story began with the isolation of large ribonucleoprotein complexes (RNPs) (Davis and Rueckert, 1972) from purified viral particles of ASLV and MoMuLV, which were able to support viral DNA synthesis. These viral RNP complexes, consisting of the 60-70S genomic RNA dimer coated by about 2000 NC molecules, represent the most stable inner structure of the viral particle, with a circular chromatin-like conformation (Chen et al, 1980;Pager et al, 1994). These viral RNPs or nucleocores also contain molecules of the viral reverse transcriptase (RT) and integrase (IN) enzymes together with molecules of cellular tRNAs and ribosomal RNA (Chen et al, 1980;Darlix et al, 1995;Dickson et al, 1985).…”

Section: Forewordsmentioning

confidence: 99%

Retrospective on the all-in-one retroviral nucleocapsid protein

et al. 2014

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This review aims at briefly presenting a retrospect on the retroviral nucleocapsid protein (NC), from an unspecific nucleic acid binding protein (NABP) to an all-in-one viral protein with multiple key functions in the early and late phases of the retrovirus replication cycle, notably reverse transcription of the genomic RNA and viral DNA integration into the host genome, and selection of the genomic RNA together with the initial steps of virus morphogenesis. In this context we will discuss the notion that NC protein has a flexible conformation and is thus a member of the growing family of intrinsically disordered proteins (IDPs) where disorder may account, at least in part, for its function as a nucleic acid (NA) chaperone and possibly as a protein chaperone vis-à-vis the viral DNA polymerase during reverse transcription. Lastly, we will briefly review the development of new anti-retroviral/AIDS compounds targeting HIV-1 NC because it represents an ideal target due to its multiple roles in the early and late phases of virus replication and its high degree of conservation.

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Electron microscopy of the nucleocapsid from disrupted Moloney murine leukemia virus and of associated type VI collagen-like filaments

Cited by 12 publications

References 54 publications

Self-assembly in vitro of purified CA-NC proteins from Rous sarcoma virus and human immunodeficiency virus type 1

Self-assembly in vitro of purified CA-NC proteins from Rous sarcoma virus and human immunodeficiency virus type 1

Distribution of minor collagens during skin development

Retrospective on the all-in-one retroviral nucleocapsid protein

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