Specific modulation of apoptosis and Bcl-xL phosphorylation in yeast by distinct mammalian protein kinase C isoforms

Saraiva, Lucı́lia; Silva, Rui D.; Pereira, Goreti; Gonçalves, Jorge; Côrte‐Real, Manuela

doi:10.1242/jcs.03033

Cited by 40 publications

(51 citation statements)

References 34 publications

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“…7A and B), decreased  m ( Fig. 8A and B) and increased mitochondrial fragmentation ( It was showed that human Bcl-xL is also a mitochondrial anti-apoptotic protein in yeast, inhibiting yeast apoptosis induced by stress stimuli such as hydrogen peroxide, menadione [27] and acetic acid [17]. Moreover, a previous work performed by our group…”

Section: Coleon U-induced Growth Inhibition In Yeast Expressing Npkc-mentioning

confidence: 73%

“…Yeast expression of a mammalian PKC isoform and/or human Bcl-xL was previously confirmed by Western blot analysis [17,19].…”

Section: Plasmidsmentioning

confidence: 92%

“…It was also showed that the yeast PKC expression system was a potential in vivo assay for the screening of PKC modulators [14,16]. Based on these data, this yeast PKC assay has been used by our group not only to study the role of PKC isoforms on apoptosis regulation [17], but also to search for isoform-selective PKC activators [18,19] and inhibitors [20,21]. This simpler eukaryotic cell model, established and validated in such studies, allows the effect of small-molecules on each PKC isoform to be analysed without the genetic complexity of the PKC family, specifically the coexistence of multiple PKC isoforms in the same mammalian cells and the extensive cross-talk amongst numerous mammalian signalling pathways.…”

Section: Page 5 Of 47mentioning

confidence: 94%

“…Constructed yeast expression plasmids YEplac181-LEU2 with the cDNA encoding for bovine PKC-, rat PKC-I or PKC-mouse PKC- or PKC-, under control of a galactose- continuous shaking, to 0.5 OD 600 (mid-log phase; about 42 h incubation), as described [17].…”

Section: Plasmidsmentioning

confidence: 99%

“…Propidium iodide (PI) and TUNEL staining to monitor plasma membrane integrity and DNA fragmentation respectively were carried out as described [17]. Briefly, about 10 7 cells incubated in galactose selective medium with 1 M coleon U or DMSO only were collected and incubated with 5 g/ml PI (Sigma-Aldrich, Sintra, Portugal) for 10 min at room for 15 min at room temperature, as described [24].…”

Section: Analysis Of Plasma Membrane Integrity Dna Fragmentation Andmentioning

confidence: 99%

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Selective activation of protein kinase C-δ and -ɛ by 6,11,12,14-tetrahydroxy-abieta-5,8,11,13-tetraene-7-one (coleon U)

Coutinho

Pereira

Simões

et al. 2009

Biochemical Pharmacology

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6,11,12,14-tetrahydroxy-abieta-5,8,11,13-tetraene-7-one (coleon U) is a diterpene compound isolated from Plectranthus grandidentatus with an antiproliferative effect on several human cancer cell lines. Herein, we studied the modulatory activity of coleon U on individual isoforms of the three protein kinase C (PKC) subfamilies, classical (cPKC- and -I), novel (nPKC- and -) and atypical (aPKC-), using a yeast PKC assay. The results showed that, whereas the PKC activator phorbol-12-myristate-13-acetate (PMA) activated every PKC tested except aPKC coleon U had no effect on aPKC and cPKCs. Besides, the effect of coleon U on nPKCs was higher than that of PMA. This revealed that coleon U was a potent and selective activator of nPKCs The isoform-selectivity of coleon U for nPKC- and - was confirmed using an in vitro PKC assay. Most importantly, while PMA activated nPKCs inducing an isoform translocation from the cytosol to the plasma membrane and a G2/M cell cycle arrest, coleon U induced nPKCs translocation to the nucleus and a metacaspase-and mitochondrial-dependent apoptosis. This work therefore reconstitutes in yeast distinct subcellular translocations of a PKC isoform and the subsequent distinct cellular responses reported for mammalian cells. Together, our study identifies a new isoform-selective PKC activator with promising pharmacological applications. Indeed, since coleon U has no effect on cPKCs and aPKC recognized as anti-apoptotic proteins, and selectively induces an apoptotic pathway dependent on nPKC- and - activation, it represents a promising compound for evaluation as an anti-cancer drug.

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Section: Coleon U-induced Growth Inhibition In Yeast Expressing Npkc-mentioning

confidence: 73%

“…Yeast expression of a mammalian PKC isoform and/or human Bcl-xL was previously confirmed by Western blot analysis [17,19].…”

Section: Plasmidsmentioning

confidence: 92%

Section: Page 5 Of 47mentioning

confidence: 94%

Section: Plasmidsmentioning

confidence: 99%

Section: Analysis Of Plasma Membrane Integrity Dna Fragmentation Andmentioning

confidence: 99%

See 3 more Smart Citations

Selective activation of protein kinase C-δ and -ɛ by 6,11,12,14-tetrahydroxy-abieta-5,8,11,13-tetraene-7-one (coleon U)

Coutinho

Pereira

Simões

et al. 2009

Biochemical Pharmacology

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Current awareness on yeast

2007

Yeast

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In order to keep subscribers up‐to‐date with the latest developments in their field, this current awareness service is provided by John Wiley & Sons and contains newly‐published material on yeasts. Each bibliography is divided into 10 sections. 1 Reviews; 2 General; 3 Biochemistry; 4 Biotechnology; 5 Cell Biology; 6 Gene Expression; 7 Genetics; 8 Physiology; 9 Medical Mycology; 10 Recombinant DNA Technology. Within each section, articles are listed in alphabetical order with respect to author. If, in the preceding period, no publications are located relevant to any one of these headings, that section will be omitted. (6 weeks journals ‐ search completed 27th. Dec. 2006)

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Tracing the Roots of Death: Apoptosis in Saccharomyces cerevisiae

Carmona-Gutiérrez

Madeo

2009

Essentials of Apoptosis

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The modulation of cell death is a complex process. Its deregulation can lead to several types of diseases ranging from cancer to neurodegenerative disorders in humans. The finding that Saccharomyces cerevisiae can also undergo apoptosis has opened doors to investigate programmed cell death in a clear-cut model organism that unifies both technical advantages and a eukaryotic ''cell room.'' So far, cell death in yeast has been described under multiple conditions, including exposure to different drugs, failure in several cellular processes, or heterologous expression of human proapoptotic genes. Yeast apoptosis has also been shown to occur in physiological scenarios such as aging or failed mating, thus suggesting a teleological explanation for the death of a unicellular organism. Finally, several yeast orthologues of crucial metazoan apoptotic regulators have been identified and characterized, including a caspase and the apoptosis-inducing factor. Uncovering apoptosis in yeast as well as in other fungi and unicellular parasites is of great medical interest. Moreover, it is helping to decipher the molecular mechanisms of cell death in higher organisms.

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Specific modulation of apoptosis and Bcl-xL phosphorylation in yeast by distinct mammalian protein kinase C isoforms

Cited by 40 publications

References 34 publications

Selective activation of protein kinase C-δ and -ɛ by 6,11,12,14-tetrahydroxy-abieta-5,8,11,13-tetraene-7-one (coleon U)

Selective activation of protein kinase C-δ and -ɛ by 6,11,12,14-tetrahydroxy-abieta-5,8,11,13-tetraene-7-one (coleon U)

Current awareness on yeast

Tracing the Roots of Death: Apoptosis in Saccharomyces cerevisiae

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