Comparison of Mitotic Phenomena and Effects Induced by Hypertonic Solutions in Hela Cells

Robbins, Elliott; Pederson, Thoru; Klein, Paul

doi:10.1083/jcb.44.2.400

Cited by 134 publications

(42 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To further confirm that these alkaloids were inhibitors of initiation, cells incubated with harringtonine were subjected to hypertonicity. Medium hypertonicity (23,24,35), itself an effective inhibitor of initiation (Fig. 1B), caused essentially complete run-off of ribosomes from RNA.…”

Section: Resultsmentioning

confidence: 94%

“…As was mentioned above, the effect of each compound on the polysome pattern was tested under three conditions: direct effect of the compound on the polysome pattern; effect on ribosome run-off produced by medium hypertonicity (23,24,35); and effect on recovery from hypertonicity. Appropriate medium hypertonicity has been shown to block initiation with subsequent run-off of ribosomes from polysomes.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Inhibition of Protein Synthesis in Intact HeLa Cells

Tscherne

Pestka

1975

Antimicrob Agents Chemother

View full text Add to dashboard Cite

Polysome analysis has proved to be a sensitive probe for the mode of action of inhibitors of protein synthesis in intact HeLa cells. To classify the active compounds as inhibitors of initiation, elongation, or termination, their effects on the cellular polyribosome pattern were compared under three conditions. These conditions tested (i) their direct effect on the polyribosome profile; (ii) their effect on ribosome run-off produced by hypertonicity; and (iii) their effects on recovery from hypertonicity. Using this technique, diacetoxyscirpenol, 2-(4-methyl-2,6-dinitroanilino)-N-methylpropionamide, and three alkaloids, harringtonine, isoharringtonine, and homoharringtonine, were found to be inhibitors of initiation. Polysome analysis indicated that in HeLa cells 7.8 x 10-7 M pactamycin, which inhibited protein synthesis 94%, interfered with elongation as well as initiation under these conditions. Emetine, anisomycin, cycloheximide, and trichodermin each gave polysome patterns consistent with inhibition of elongation. Fusidic acid and aurintricarboxylic acid inhibited incorporation of ["CC]

show abstract

Section: Resultsmentioning

confidence: 94%

mentioning

confidence: 99%

Inhibition of Protein Synthesis in Intact HeLa Cells

Tscherne

Pestka

1975

Antimicrob Agents Chemother

View full text Add to dashboard Cite

show abstract

“…As demonstrated by Robbins et al (Robbins et al, 1970) and Pederson and Robbins (Pederson and Robbins, 1970), hypertonic shock does not create mitotic chromatin condensation. Concordantly, we did not observe individual chromosomes or mitotic banding.…”

Section: Does Molecular Crowding Affect Nuclear Architecture?mentioning

confidence: 92%

Experimental evidence for the influence of molecular crowding on nuclear architecture

Richter

Neßling

Lichter

2007

Journal of Cell Science

112

124

View full text Add to dashboard Cite

Many compounds in the cell nucleus are structurally organized. To assess the influence of structural organization on nuclear function, we investigated the physical mechanisms of structure formation by using molecular crowding as a parameter for nuclear integrity. Molecular crowding promotes compaction of macromolecular compounds depending on their size and shape without the need for site-specific interactions. HeLa and MCF7 cells were incubated with hypertonic medium to increase crowding of their macromolecular content as a result of the osmotic loss of water. Supplementation of sucrose, sorbitol or NaCl to the growth medium shifted nuclear organization, observed by fluorescence and electron microscopy, towards compaction of chromatin and segregation of other nuclear compounds. With increasing hypertonic load and incubation time, this nuclear re-organization proceeded gradually, irrespective of the substances used, and reversibly relaxed to a regular phenotype upon re-incubation of cells in isotonic growth medium. Gradual and reversible re-organization are major features of controlled de-mixing by molecular crowding. Of fundamental importance for nuclear function, we discuss how macromolecular crowding could account for the stabilization of processes that involve large, macromolecular machines.

show abstract

“…The cellular effects of changes in tonicity have been investigated extensively (Robbins et al, 1970;Banuett, 1998;Gustin et al, 1998;Lang et al, 1998). In particular, analysis of the yeast Saccharomyces cerevisiae has elucidated major roles for two MAP kinase signaling pathways: the Pkc1p "cell integrity" pathway, which is stimulated by hypotonic shock (Watanabe et al, 1994;Davenport et al, 1995), and the Hog1p pathway, which is stimulated by hypertonic shock (Brewster et al, 1993;Posas and Saito, 1997).…”

Section: Introductionmentioning

confidence: 99%

Perturbation of the Nucleus: A Novel Hog1p-independent, Pkc1p-dependent Consequence of Hypertonic Shock in Yeast

Nanduri

Tartakoff

2001

MBoC

View full text Add to dashboard Cite

Hypertonic shock of Saccharomyces cerevisiae activates the Hog1p MAP kinase cascade. In contrast, protein kinase C (Pkc1p) and the "cell integrity" MAP kinase cascade are critical for the response to hypotonic shock. We observed that hypertonic shock transiently relocated many, but not all, nuclear and nucleolar proteins to the cytoplasm. We hypothesized that the relocation of nuclear proteins was due to activation of the Hog1p kinase cascade, yet, surprisingly, Hog1p was not required for these effects. In contrast, Pkc1p kinase activity was required, although the Pkc1p MAP kinase cascade and several factors known to lie upstream and downstream of Pkc1p were not. Moreover, sudden induction of a hyperactive form of Pkc1p was sufficient to relocate nuclear proteins. Taken together, these observations show that the scope of involvement of Pkc1p in the organization of the nucleus considerably exceeds what has been characterized previously. The relocation of nuclear proteins is likely to account for the profound inhibition of RNA synthesis that was observed during hypertonic shock. INTRODUCTIONThe cellular effects of changes in tonicity have been investigated extensively (Robbins et al., 1970;Banuett, 1998;Gustin et al., 1998;Lang et al., 1998). In particular, analysis of the yeast Saccharomyces cerevisiae has elucidated major roles for two MAP kinase signaling pathways: the Pkc1p "cell integrity" pathway, which is stimulated by hypotonic shock (Watanabe et al., 1994;Davenport et al., 1995), and the Hog1p pathway, which is stimulated by hypertonic shock (Brewster et al., 1993;Posas and Saito, 1997). Each pathway is required for long-term survival in the corresponding medium. Hypertonic shock profoundly affects the actin cytoskeleton (Chowdhury et al., 1992;Mulholland et al., 1994;Delley and Hall, 1999).The Pkc1p pathway can receive input from plasma membrane glycoproteins of the Wsc family and from the Rho GTPases and their regulatory factors, for example in the context of control of cell polarization and cell wall biosynthesis (Gustin et al., 1998). Interestingly, some signaling events that require Pkc1p appear not to require the corresponding downstream MAP kinase cascade, which is linked to activation of the transcription factors Rlm1p and Swi4p/ Swi6p (Lee and Levin, 1992;Gustin et al., 1998;Delley and Hall, 1999; Li et al., 2000). In contrast, the Hog1p pathway is initiated by a two-component phosphorelay involving the cell surface transmembrane protein Sln1p and the cytosolic protein Ypd1p (Posas et al., 1996). Stimulation of this pathway causes phosphorylation and transient nuclear entry of the terminal kinase, Hog1p, as well as of the transcription factor Msn2p (Ferrigno et al., 1998;Gö rner et al., 1998;Reiser et al., 1999). The downstream substrate(s) of Hog1p kinase is not known.We have observed recently that arrest of the secretory pathway in yeast inhibits nuclear import and causes many nucleolar and nucleoplasmic proteins to relocate to the cytoplasm. These events (the "arrest of secretion response") a...

show abstract

Comparison of Mitotic Phenomena and Effects Induced by Hypertonic Solutions in Hela Cells

Cited by 134 publications

References 18 publications

Inhibition of Protein Synthesis in Intact HeLa Cells

Inhibition of Protein Synthesis in Intact HeLa Cells

Experimental evidence for the influence of molecular crowding on nuclear architecture

Perturbation of the Nucleus: A Novel Hog1p-independent, Pkc1p-dependent Consequence of Hypertonic Shock in Yeast

Contact Info

Product

Resources

About