Replication of rous sarcoma virus and the biosynthesis of the oncogenic subviral ribonucleoprotein particles (“virosomes”) in the mitochondria isolated from rous sarcoma tissue

Kára, J.; Mach, O; Cerná, H

doi:10.1016/s0006-291x(71)80173-0

Cited by 52 publications

(10 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Finally, it is also possible that the transformation-associated change in mitochondrial distribution observed here may reflect other biochemical and functional changes that may occur in the mitochondria of transformed cells (21). In accordance with such a possibility is the report that Rous sarcoma virus RNA and Rous sarcoma virus associated proteins are present in the mitochondria of cells infected with Rous sarcoma virus (38). Rat …”

Section: Discussionsupporting

confidence: 78%

Localization of mitochondria in living cells with rhodamine 123.

Johnson¹,

Walsh²,

Chen³

1980

Proc. Natl. Acad. Sci. U.S.A.

1,372

721

View full text Add to dashboard Cite

The laser dye rhodamine 123 is shown to be a specific probe, for the localization of mitochondria in living cells.By virtue of its selectivity for mitochondria and its fluorescent properties, the detectability of mitochondria stained with rhodamine 123 is significantly improved over that provided by conventional light microscopic techniques. With the use of rhodamine 123, it is possible to detect alterations in mitochondrial distribution following transformation by Rous sarcoma virus and changes in the shape and organization of mitochondria induced by colchicine treatment.Since the classical investigations of the. mitochondrion some 90 years ago, much work has been carried out on the structure and function of this complex organelle. Extensive biochemical studies of mitochondria have proved that they play a cardinal role in the generation of energy essential for the survival and proliferation of eukaryotic cells (1-4). As intracellular organelles, mitochondria show remarkable plasticity, mobility, and morphological heterogeneity (5-13). Mitochondrial morphology is influenced by the metabolic state of the cell, cell cycle, cellular development and differentiation, and by pathological states (14)(15)(16)(17)(18)(19)(20). In addition, both morphological and functional changes in mitochondria have been shown to occur in conjunction with neoplastic transformation (21).Although isolated mitochondria and mitochondria in fixed cells have been extensively investigated, much less attention has been paid to mitochondria in living cells. Previous investigations of mitochondria in living cells have been hampered by the lack of techniques allowing high-resolution visualization of these organelles. Use of Janus Green B, a dye relatively specific for mitochondria, aids somewhat in their recognition but also causes mitochondrial distortion and cytotoxic effects (5,22 sponsible for mitochondrion-specific staining in such a preparation has been characterized as rhodamine 3B (unpublished data). This finding prompted us to screen various rhodamine compounds for mitochondrion-specific staining. The results reported here describe the use of the laser dye rhodamine 123 as a specific fluorescent probe for mitochondria in living cells. Rhodamine 123 stains mitochondria directly (without passage through endocytic vesicles and lysosomes) and provides lowbackground high-resolution fluorescent images of mitochondria without apparent cytotoxic effects. MATERIALS AND METHODSThe purified laser dye rhodamine 123 (Eastman) was dissolved in double-distilled water at a concentration of 1 mg/ml and subsequently diluted to 10 ,g/ml in Dulbecco's modified Eagle's medium (GIBCO). Cultured cells grown on 12-mm round glass coverslips (Rochester Scientific, Rochester, NY) were incubated with rhodamine 123 (10 ,ug/ml) for 30 min in a 10% CO2 incubator at 37°C. Coverslips were then rinsed through three 5-ml changes of medium (5 min per rinse) and mounted in medium supplemented with 10% fetal calf serum (GIBCO) on a live-cell observation chamber prep...

show abstract

Section: Discussionsupporting

confidence: 78%

Localization of mitochondria in living cells with rhodamine 123.

Johnson¹,

Walsh²,

Chen³

1980

Proc. Natl. Acad. Sci. U.S.A.

1,372

721

View full text Add to dashboard Cite

show abstract

“…1). It can be excluded that the mitochondrial y-polymerase represents an RNA-dependent DNA polymerase of a murine C-type virus in our cultures [21,22] because of (a) its inability to copy a natural RNA template (Table 1) [23,24], (b) its molecular The DNA polymerase was solubilized from isolated organelles as described in Materials and Methods. The dialysate was adsorbed to a 2-ml DEAE-cellulose column.…”

Section: Identification Of the Mitochondria1 Dna Polymerase From Mousmentioning

confidence: 99%

Changes of Mitochondrial DNA Polymerase-gamma Activity in Synchronized Mouse Cell Cultures

Radsak

Schütz

1978

Eur J Biochem

View full text Add to dashboard Cite

Following synchronization by a double hydroxyurea block, mouse cell cultures exhibited a period of accelerated precursor incorporation into mitochondrial DNA during the late nuclear S phase. Peak activity of mitochondrial DNA polymerase-y occurred concurrent to the interval of accelerated organelle DNA synthesis. Mixing experiments suggested that the variations in mitochondrial DNA polymerase activity during the cell cycle were not due to free inhibitors in the enzyme preparations examined.There is well-documented evidence that mammalian cells possess in addition to the DNA polymerasesa, / 3 and y, a DNA polymerase activity associated with the mitochondria [l]. Recent reports on the characterization of this organelle DNA polymerase were, however, inconsistent with respect to the properties of the enzyme preparations described [2,3]. In contrast to earlier investigations by Kalf and Ch'ih [4] as well as Meyer and Simpson [5] and a recent report by Hecht [3] which deal with mitochondrial DNA polymerase from liver cells, the organelle enzyme from HeLa cells [2,6] did not exhibit a significant salt dependence and ethidium bromide sensitivity. Furthermore, a chloramphenicol treatment of HeLa cells was found to result in a considerable loss of this enzyme activity [7]. The latter observation could be interpreted in favour of the suspicion that the HeLa cell mitochondrial DNA polymerase is a procaryotic enzyme originating, for instance, from a mycoplasma contamination not detectable by routine examinations [7,8]. Attempts from this laboratory succeeded in isolating a DNA polymerase activity from a mitochondrial replication complex of mouse cells [8] which was most likely free of exogenous contaminations as a result of the isolation procedure. More than 98% of the activity solubilized from this complex showed distinct properties identical to those of the enzyme described by Hecht [3] purpose of this investigation was to look for a correlation between mitochondrial DNA synthesizing activity and the mitochondrial DNA polymerase activity during the cell cycle of synchronized mouse cells in order to further establish the function of the enzyme. It will be shown that there is a period of accelerated precursor incorporation into mitochondrial DNA [lo] during the cell cycle which is paralleled by the activity peak of the mitochondrial DNA polymerase-y. MATERIALS AND METHODS CellsCl-1D cells, a permanent mouse cell line [11,12] which is deficient in the major cellular thymidine kinase activity, although not deficient in the mitochondrial thymidine kinase, were used for all experiments. The monolayer cultures were cultivated in F-16 medium (Gibco) supplemented with 5 % calf serum, 200 units penicillin/ml and 10 pg 5-bromodeoxyuridine/ml. This cell line incorporates exogenous thymidine mainly into mitochondrial DNA and cannot replicate in medium containing hypoxanthine (1 5 pg/ ml), aminopterin (20 pg/ml) and thymidine (4 pg/ml). Routine examinations for mycoplasma contamination were found to be negative. Synchronization Pro...

show abstract

“…Moreover, the virus uses ATP for its own benefit. The anti-uncoupling effect was, in part, previously observed with nonpurified virus [7], Recent studies furnish proof of a close correlation between the replication of some viruses with mitochondrial respiration [14,15] localized within the inner membrane [16]. More precisely, the RNA of Venezuelan equine encephalomyelitis virus (a togavirus like rubella virus) can penetrate isolated rat liver mitochondria [17], It has also been shown that virus replication occurred within the mitochondria [18] and that upon infec tion, the mitochondria switched to production of virus-specified products as opposed to mitochondrial-specific products [19].…”

Section: Resultsmentioning

confidence: 60%

Respiration and ATP Level in BHK21/13S Cells during the Earliest Stages of Rubella Virus Replication

Bardeletti¹

1977

Intervirology

View full text Add to dashboard Cite

This study is concerned with the effect of virus infection on the energy balance of the cell and the possible involvement of mitochondria in virus replication. The process of rubella virus adsorption and penetration is immediately associated with a stimulation of O₂ uptake. Shortly afterwards, ATP levels decline, possibly because of increased ATP utilization. Virus infection appears to protect mitochondria against the uncoupling effect of dinitrophenol.

show abstract

Replication of rous sarcoma virus and the biosynthesis of the oncogenic subviral ribonucleoprotein particles (“virosomes”) in the mitochondria isolated from rous sarcoma tissue

Cited by 52 publications

References 15 publications

Localization of mitochondria in living cells with rhodamine 123.

Localization of mitochondria in living cells with rhodamine 123.

Changes of Mitochondrial DNA Polymerase-gamma Activity in Synchronized Mouse Cell Cultures

Respiration and ATP Level in BHK21/13S Cells during the Earliest Stages of Rubella Virus Replication

Contact Info

Product

Resources

About