On the association of mRNA with the cytoskeleton in uninfected and adenovirus-infected human KB cells

Venrooij, Walther J. van; Sillekens, Peter; Eekelen, Chris A.G. van; Reinders, Rita J.

doi:10.1016/0014-4827(81)90301-3

Cited by 140 publications

(89 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It is estimated that 70 -80% of mRNA in the cell is associated with, but not bound directly to, microfilaments or microtubules (Lenk et al, 1977;Cervera et al, 1981;van Venrooij et al, 1981;Taneja et al, 1992;Bassell, 1993;Bassell et al, 1994). A linker is therefore required for the association of mRNAs to the cytoskeleton.…”

Section: Introductionmentioning

confidence: 99%

Interactions of Elongation Factor 1α with F-Actin and β-Actin mRNA: Implications for Anchoring mRNA in Cell Protrusions

Liu

Grant²,

Persky³

et al. 2002

MBoC

160

159

View full text Add to dashboard Cite

The targeting of mRNA and local protein synthesis is important for the generation and maintenance of cell polarity. As part of the translational machinery as well as an actin/ microtubule-binding protein, elongation factor 1␣ (EF1␣) is a candidate linker between the protein translation apparatus and the cytoskeleton. We demonstrate in this work that EF1␣ colocalizes with ␤-actin mRNA and F-actin in protrusions of chicken embryo fibroblasts and binds directly to F-actin and ␤-actin mRNA simultaneously in vitro in actin cosedimentation and enzyme-linked immunosorbent assays. To investigate the role of EF1␣ in mRNA targeting, we mapped the two actin-binding sites on EF1␣ at high resolution and defined one site at the N-terminal 49 residues of domain I and the other at the C-terminal 54 residues of domain III. In vitro actin-binding assays and localization in vivo of recombinant full-length EF1␣ and its various truncates demonstrated that the C terminus of domain III was the dominant actin-binding site both in vitro and in vivo. We propose that the EF1␣-F-actin complex is the scaffold that is important for ␤-actin mRNA anchoring. Disruption of this complex would lead to delocalization of the mRNA. This hypothesis was tested by using two dominant negative polypeptides: the actin-binding domain III of EF1␣ and the EF1␣-binding site of yeast Bni1p, a protein that inhibits EF1␣ binding to F-actin and also is required for yeast mRNA localization. We demonstrate that either domain III of EF1␣ or the EF1␣-binding site of Bni1p inhibits EF1␣ binding to ␤-actin mRNA in vitro and causes delocalization of ␤-actin mRNA in chicken embryo fibroblasts. Taken together, these results implicate EF1␣ in the anchoring of ␤-actin mRNA to the protrusion in crawling cells.

show abstract

Section: Introductionmentioning

confidence: 99%

Interactions of Elongation Factor 1α with F-Actin and β-Actin mRNA: Implications for Anchoring mRNA in Cell Protrusions

Liu

Grant²,

Persky³

et al. 2002

MBoC

160

159

View full text Add to dashboard Cite

show abstract

“…This distribution can undergo substantial alteration with changing extracellular milieu [l]. Recent evidence (reviewed in [2]) has suggested that a proportion or possibly all of the 'free' polysomes are in some way associated with the cytoskeleton [3,4]. The evidence comes both from electron microscopy and from the observed retention of polysomes in the insoluble cytoskeletal material obtained from cells treated with a nonionic detergent.…”

Section: Introductionmentioning

confidence: 99%

“…The evidence comes both from electron microscopy and from the observed retention of polysomes in the insoluble cytoskeletal material obtained from cells treated with a nonionic detergent. Such polysomes are retained in the cytoskeleton in the presence of 0.05% non-ionic detergent, but are released upon the addition of 1% deoxycholate [3,5]. This method is, however, very similar to that used to prepare membranebound polysomes [6,7], so that it would seem imprecise to define the cytoskeletal-bound polysomes…”

Section: Introductionmentioning

confidence: 99%

Evidence that insulin increases the proportion of polysomes that are bound to the cytoskeleton in 3T3 fibroblasts

Hesketh¹,

Pryme

1988

FEBS Letters

View full text Add to dashboard Cite

The association of polysome redistribution with changes in protein synthesis was investigated in insulin-stimulated fibroblasts. Free polysomes were released by Nonidet-P40 and 25 mM KCI, cytoskeletal-bound polysomes were retained at 25 mM KC1 but released at 130 mM, while membrane-bound polysomes were released by deoxycholate. Insulin increased the proportion of polysomes which were retained at 25 mM KC1 but had no effect when extraction was carried out at 130 mM KCl, suggesting that more polysomes were associated with the cytoskeleton. Insulin also reduced the amount of actin released from the detergent-insoluble cytoskeleton indicating that the hormone affects microfilament organization.

show abstract

“…Gentle disruption of eukaryotic cells with nonionic detergents permits concomitant biochemical and morphological analyses of the CF after differential centrifugation is used to separate the cell lysate into detergent soluble and insoluble fractions. An important result of these biochemical studies is that most of the polyribosomal mRNA is found to be associated with the CF (8,20,33).The purpose of the present investigation was to test the possibility of mRNA interaction with the CF by cytological methods, in particular in situ hybridization. In this paper, in situ hybridization with poly(U) or a cloned actin DNA probe and radioactive isotope tracing methods have been used to demonstrate that a significant proportion of the mRNA of follicle ceils is associated with the CF.…”

mentioning

confidence: 98%

mentioning

confidence: 99%

Messenger RNA in the cytoskeletal framework: analysis by in situ hybridization.

Jeffery¹

1982

The Journal of Cell Biology

View full text Add to dashboard Cite

The possibility of an association of mRNA with the cytoskeletal framework (CF) of ascidian (Styela plicata) follicle cells was examined in this study. The approach was to extract the follicle cells with Triton X-100 and determine whether mRNA persisted in the insoluble residue by two methods, in situ hybridization with poly(U) and actin DNA probes and the incorporation of radioactive isotopes into RNA. Triton X-100 extraction of follicle cells yielded a filamentous CF containing ~70% of the total poly (A) but only 9% of the total lipid, 23% of the total protein, and 28% of the total RNA. In situ hybridization with a poly (U) probe indicated that ~70% of the poly (A) was associated with the CF. In situ hybridization with a cloned actin DNA probe indicated that ~60% of the actin mRNA was associated with the CF. Autoradiography of detergent-extracted follicle cells, which had been labeled with [3H]uridine or [3H]adenosine, indicated that >90% of the newly synthesized poly (A)+RNA was preserved in the CF. Thus more newly synthesized mRNA than steady-state mRNA may be present in the Triton X-100 insoluble fraction. It is concluded that a significant proportion of the mRNA complement of ascidian follicle cells is associated with the CF.The cytoplasm of many eukaryotic cells contains an elaborate network of filaments and associated components which has been termed the cytoskeletal framework (CF) (21). The CF includes microtubules, microfdaments, intermediate filaments, microtrabeculae (36), centrioles (20), a membrane protein lamina (2), and probably many other cellular components. It can be visualized by immunofluorescence microscopy of intact cells (19,24,25,35 ) or by electron microscopy of detergent-extracted cells (3,20,27,29,30). Gentle disruption of eukaryotic cells with nonionic detergents permits concomitant biochemical and morphological analyses of the CF after differential centrifugation is used to separate the cell lysate into detergent soluble and insoluble fractions. An important result of these biochemical studies is that most of the polyribosomal mRNA is found to be associated with the CF (8,20,33).The purpose of the present investigation was to test the possibility of mRNA interaction with the CF by cytological methods, in particular in situ hybridization. In this paper, in situ hybridization with poly(U) or a cloned actin DNA probe and radioactive isotope tracing methods have been used to demonstrate that a significant proportion of the mRNA of follicle ceils is associated with the CF. The follicle cells of ascidian eggs were selected as a model system because they are known to contain a filamentous cytoskeleton (9, 10) and are extremely active in the synthesis and accumulation of THt IOURNA[ OE CELL 81OtOGY • VOLUME 95 OCTOBER 1982 1-7 © The Rockefeller University Press -0021-9525/82/10/1/7 $1.00 poly(A)+RNA during oogenesis and early development (15). The association of mRNA with the CF may be a mechanism for controlling the polarity of mRNA distribution in eukaryotic cells. MATERIALS AND M...

show abstract

On the association of mRNA with the cytoskeleton in uninfected and adenovirus-infected human KB cells

Cited by 140 publications

References 23 publications

Interactions of Elongation Factor 1α with F-Actin and β-Actin mRNA: Implications for Anchoring mRNA in Cell Protrusions

Interactions of Elongation Factor 1α with F-Actin and β-Actin mRNA: Implications for Anchoring mRNA in Cell Protrusions

Evidence that insulin increases the proportion of polysomes that are bound to the cytoskeleton in 3T3 fibroblasts

Messenger RNA in the cytoskeletal framework: analysis by in situ hybridization.

Contact Info

Product

Resources

About