The metabolism of SV40 RNA is associated with the cytoskeletal framework

Ben-Ze’ev, Avri; Horowitz, Mia; Skolnik, H; Abulafia, Rivka; Laub, Orgad; Aloni, Yosef

doi:10.1016/0042-6822(81)90350-0

Cited by 54 publications

(29 citation statements)

References 25 publications

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“…Although the detailed mechanism by which cellular polysomes are bound to this structure is still obscure, Penman and co-workers extended their conclusion to the case of poliovirus [6] and cells infected with vesicular stomatitis virus [5]: the viral inRNA involved in translation was found associated with the cytoskeleton. This conclusion was recently confirmed for the case of KB cells infected with adenovirus [ 7 ] and for the BSC-1 cells infected with SV40 [8]. The general conclusion of these studies is that no unique class of cellular or viral mRNA is translated while bound to the cytoskeleton, with one exception [9].…”

supporting

confidence: 64%

“…1). This apparent discrepancy results from the fact that only the most stable cellular mRNAs are analysed but confirms the previous observation that 'old' poly(A)-rich RNA, not engaged in polysomes, accumulate in the soluble fraction [5,8]. In contrast, a higher proportion of this poly(A)-rich RNA is found in the cytoskeletal fraction from vaccinia-virus-infected cells (Table 6).…”

Section: Fate Of Cellular Poly(a)-rich R N a In Eat Cells Infected Wimentioning

confidence: 38%

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Expression of Vaccinia Virus Early mRNA in Ehrlich Ascites Tumor Cells

Lemieux

Beaud

1982

European Journal of Biochemistry

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A method for preparing detergent cytoskeletons from uninfected or vaccinia-virus-infected cells is described. This method resulted in the fractionation of the cytoplasmic compartment into soluble and cytoskeletal fractions. More than 85% of small molecules and tRNA were released from the cytoskeleton and recovered into the soluble fraction. The cytoskeletal fraction contained about 40% of the cytoplasmic proteins and 6?':,, of total cytoplasmic RNA. Similar values were obtained for vaccinia-virus-infected cells. In contrast, whereas 85 ?,, of the cellular polysomes and poly(A)-rich RNA remained associated with the cytoskeleton in uninfected cells, more than 40% of vaccinia early mRNA engaged into polysomes was found to be not associated with the cytoskeleton. A similar partition was found when the viral R N A was labeled for 15 min at 5 min and 20 min after the infection or when varying the duration of the pulse. Soluble and cytoskeletal viral polysomes were found to synthesize a similar set of proteins after translation in vitro of the corresponding mRNA. The fate of rapidly labeled cellular poly(A)-rich RNA upon vaccinia virus infection was followed by a glucosaminei uridine chase procedure, and also that of relatively stable poly(A)-rich RNA after long-term labeling and chase. In both cases no release of poly(A)-rich RNA from the cytoskeleton occurred after vaccinia virus infection. These experiments reveal that cellular mRNA remains associated to the cytoskeleton in EAT cells infected with vaccinia virus (early period) whereas at least 40% of the vaccinia early polysomes are not associated with the cytoskeleton. A model for vaccinia early mRNA metabolism is presented, which may account for the rapid shut-off of host protein synthesis.The cytoplasm of animal cells contains several types of protein filaments constituting the cytoskeleton. These include the microtubules, intermediate and microfilaments and also a microtrabecular lattice [I] although the existence of the latter is questioned [2]. One important observation of these electron microscopy studies was that the so-called free polysomes (i.e. not membrane-bound) were indeed seen as being bound to the microtrabecular lattice [3]. This observation was supported by a biochemical analysis of the detergent cytoskeletons prepared from HeLa cells: it was concluded that messenger is translated only when bound to the cytoskeletal framework [4,5]. Although the detailed mechanism by which cellular polysomes are bound to this structure is still obscure, Penman and co-workers extended their conclusion to the case of poliovirus [6] and cells infected with vesicular stomatitis virus [5]: the viral inRNA involved in translation was found associated with the cytoskeleton. This conclusion was recently confirmed for the case of KB cells infected with adenovirus [ 7 ] and for the BSC-1 cells infected with SV40 [8]. The general conclusion of these studies is that no unique class of cellular or viral mRNA is translated while bound to the cytoskeleton, with one exception [9]...

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supporting

confidence: 64%

Section: Fate Of Cellular Poly(a)-rich R N a In Eat Cells Infected Wimentioning

confidence: 38%

Expression of Vaccinia Virus Early mRNA in Ehrlich Ascites Tumor Cells

Lemieux

Beaud

1982

European Journal of Biochemistry

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“…The term "skeletal framework" was suggested to include the entire structure obtained after detergent extraction of the intact cell. It consists of the cytoplasmic filaments, the plasma lamina, and possibly the nuclear matrix (6,10). This report is a study of the association of glycoconjugates with the skeletal framework.…”

mentioning

confidence: 99%

Association of glycoconjugates with the cytoskeletal framework.

Ben-Ze’ev¹,

Abulafia²

1983

Mol. Cell. Biol.

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The association of glycoconjugates with the cytoskeletal framework was examined in detergent-extracted cells. Sparse cultures offibroblasts that assemble only minimal amounts of extracellular matrix were extracted under mild conditions with Triton X-100 which remove most of the lipids and soluble cellular proteins. The detergent-resistant framework retains lectin binding sites in the nucleus, in the perinuclear area occupied by the rough endoplasmic reticulumGolgi system of the intact cell, and in a network throughout the cytoskeletal framework. Fluorescent-antibody staining with antibody against collagen type I and fibronectin reveals extensive perinuclear staining of the remnant rough endoplasmic reticulum-Golgi system. In contrast, only sporadic staining of the pericellular area is obtained with these antibodies, in sparse cultures of whole cells. Lectin binding sites were detected in the nucleus and are attributed to chromatin-associated glycoconjugates. They can be removed by DNase under conditions that preserve the cytoplasmic lectin binding sites and the nuclear matrix. The results suggest a high degree of integration of the membrane residues of the cytoplasmic elements and the nuclear matrix with the skeletal framework and indicate a possible role for the glycoconjulgates in this structural integration.The study of subcellular structure and its organization indicates the existence of highly interconnected filamentous networks in the cytoplasm of eucaryotic cells. These include the major filaments, i.e., microfilaments, intermediate filaments, and microtubules, which are also termed "cytoskeleton." A more detailed insight into the organization of the cytoplasmic networks is afforded by using a high-voltage electron microscope to examine critical point-dried whole cells (8,22,23). These studies reveal a dense, highly interconnected three-dimensional network, termed "microtrabeculae" by Wolosewick and Porter (22,23) as Triton X-100, and several laboratories have used these "Triton skeletons" for biochemical and morphological studies (4, 6, 7, 9, 10, 14, 18, 21). This extraction removes most of the lipids and the soluble cellular proteins, leaving intact the structural elements, and reveals aspects of cell structure such as a lamina that forms the outer boundary of the extracted cell (4). The term "skeletal framework" was suggested to include the entire structure obtained after detergent extraction of the intact cell. It consists of the cytoplasmic filaments, the plasma lamina, and possibly the nuclear matrix (6,10). This report is a study of the association of glycoconjugates with the skeletal framework.The results suggest that glycoconjugates and the proteinaceous membrane moieties of the rough endoplasmic reticulum (RER)-Golgi system are intimately associated with the skeletal framework. The nucleus also contains glycoconjugates which can be removed by dissolution of the chromatin. The extraction of chromatin leaves the cytoskeletal lectin binding sites intact together with a nuclear matrix. These studies...

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“…Our studies revealed an association with the cytoskeleton of the nuclear viruses simian virus 40 (SV40) and herpes simplex virus (HSV) at various stages in replication (1,(5)(6)(7)10). We found that the biogenesis, transport, and activity of SV40 mRNA are associated with the nuclear matrix and the cytoskeleton (1,5,7,10) and that the transport of viral proteins from the cytoplasm to the nucleus and the assembly of viruses occur in association with cellular networks (6). However, despite extensive information about the molecular biology of viral replication, very little is known about the mechanisms by which viruses damage the infected cells.…”

mentioning

confidence: 99%

“…1, 2, 3, and 7. DISCUSSION Recent studies have demonstrated that various steps in the replication of SV40 are associated with the subcellular cytoskeletal network and with the nuclear matrix (1,5,7,10,24,35). In this report we demonstrated that at late times after SV40 infection, there is an alteration in the organization of the vimentin-containing IF network which is accompanied by a decrease in vimentin synthesis.…”

mentioning

confidence: 99%

Inhibition of vimentin synthesis and disruption of intermediate filaments in simian virus 40-infected monkey kidney cells.

Ben-Ze’ev¹

1984

Mol. Cell. Biol.

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The organization, synthesis, and phosphorylation of vimentin were studied at various times after infection of monkey kidney cells with simian virus 40. Late after infection (between 36 and 48 h postinfection) there is a dramatic reduction in vimentin synthesis that is paralleled by a specific disruption of the intermediate filament network. At the same time there is no apparent alteration of the organization or the synthesis of the actincontaining filaments and of the microtubules. The inhibition of vimentin synthesis is also reflected by the level of vimentin mRNA activity in the infected cells, as assayed in a cell-free in vitro translation system, and vimentin mRNA concentration as revealed by RNA blot hybridization to cloned vimentin cDNA. The level of vimentin phosphorylation also decreases dramatically but at a much earlier time after infection (between 14 and 24 h postinfection), when mitosis in the infected cells is blocked. Although the decrease in vimentin synthesis in simian virus 40-infected cells is paralleled by the alterations in the organization of the intermediate filament network, the phosphorylation of vimentin correlates with the cell cycle, as it does in other systems. A possible feedback control mechanism of vimentin synthesis by alterations in the organization of the intermediate filament network is discussed.Studies of cell structure have demonstrated the existence of a proteinaceous cytoskeletal network that has been implicated primarily in fulfilling structural roles, such as determining cell shape and cell locomotion and the integration and positioning of the cellular oganelles (for a review see reference 14). Increasing interest in the filamentous networks of the cell was raised by studies suggesting a correlation between changes in cell shape and the control of growthrelated phenomena (2, 3, 8, 9, 19; for a review see A. BenZe'ev, in J. W. Shay, ed., Cell and Muiscle Motility, vol. 6, in press).In addition, numerous studies have demonstrated an association between the replication of a wide variety of viruses and the cytoskeletal network of the infected cell (for a review see reference 29). As a model system for investigating the possible functional roles of the complex cellular networks, we used nuclear DNA virus-infected cells and followed the metabolism of viral gene products in relation to the cytoskeleton. Our studies revealed an association with the cytoskeleton of the nuclear viruses simian virus 40 (SV40) and herpes simplex virus (HSV) at various stages in replication (1,(5)(6)(7)10). We found that the biogenesis, transport, and activity of SV40 mRNA are associated with the nuclear matrix and the cytoskeleton (1,5,7,10) and that the transport of viral proteins from the cytoplasm to the nucleus and the assembly of viruses occur in association with cellular networks (6). However, despite extensive information about the molecular biology of viral replication, very little is known about the mechanisms by which viruses damage the infected cells. In the present study we Cell f...

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The metabolism of SV40 RNA is associated with the cytoskeletal framework

Cited by 54 publications

References 25 publications

Expression of Vaccinia Virus Early mRNA in Ehrlich Ascites Tumor Cells

Expression of Vaccinia Virus Early mRNA in Ehrlich Ascites Tumor Cells

Association of glycoconjugates with the cytoskeletal framework.

Inhibition of vimentin synthesis and disruption of intermediate filaments in simian virus 40-infected monkey kidney cells.

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