Characterization and Determination of Cell Death by Apoptosis

Reynolds, Jason E.; McBain, John A.; Zhou, Ping; Eastman, Alan; Craig, Ruth W.

doi:10.1002/0471250570.spi050

Cited by 8 publications

(15 citation statements)

References 32 publications

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“…Differential MCL1 phosphorylation by TPA okadaic acid taxol AM Domina et al purpose, and we had previously successfully transfected these cells with MCL1 (Reynolds et al, 1994(Reynolds et al, , 1996. In addition, because our antibodies detect the human but not mouse or Chinese hamster MCL1 (i.e., no bands are seen on 1D Western analysis of CHO cells (Reynolds et al, 1994)), the transfected human MCL1 gene product can be monitored without interference from endogenous Chinese hamster MCL1. We thus transiently transfected CHO cells with a wild-type MCL1 construct and carried out 2D-phosphopeptide mapping.…”

Section: Resultsmentioning

confidence: 98%

“…The cell lines used in these experiments include the BL41-3 Burkitt lymphoma subline (Vrana et al, 2002), in which the conditions for inducing MCL1 phosphorylation with various agents have been well established (Domina et al, 2000), and the 5AHSMyc CHO cell line (Reynolds et al, 1994(Reynolds et al, , 1996, used as a readily transfectable system for testing MCL1 mutant constructs. Previously described MCL1-transfected FDC-P1 cells (FDC-P1-S3 cells, which constitutively express MCL1 under the control of the CMV promoter, and FDC-P1-7.5 cells that express MCL1 under the control of dexamethasone-inducible pMAM promoter; Zhou et al, 1997) were used for experiments with stably transfected cells.…”

Section: Cell Culture and Drug Treatmentmentioning

confidence: 99%

“…Our laboratory has focused on another antiapoptotic member of the BCL2 family, MCL1 (Kozopas et al, 1993;Reynolds et al, 1994Reynolds et al, , 1996Zhou et al, 1997Zhou et al, , 1998Zhou et al, , 2001. Initial studies carried out in human myeloblastic leukemia (ML-1) cells demonstrated that TPA stimulates a rapid increase in MCL1 mRNA expression prior to the onset of monocytic differentiation (Kozopas et al, 1993;Yang et al, 1996;Townsend et al, 1998Townsend et al, , 1999.…”

Section: Introductionmentioning

confidence: 99%

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MCL1 is phosphorylated in the PEST region and stabilized upon ERK activation in viable cells, and at additional sites with cytotoxic okadaic acid or taxol

et al. 2004

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BCL2 family members are subject to regulation at multiple levels, providing checks on their ability to contribute to tumorigenesis. However, findings on posttranslational BCL2 phosphorylation in different systems have been difficult to integrate. Another antiapoptotic family member, MCL1, exhibits a difference in electrophoretic mobility upon phosphorylation induced by an activator of PKC (12-O-tetradecanoylphorbol 13-acetate; TPA) versus agents that act on microtubules or protein phosphatases 1/2A. A multiple pathway model is now presented, which demonstrates that MCL1 can undergo distinct phosphorylation events -mediated through separate signaling processes and involving different target sites -in cells that remain viable in the presence of TPA versus cells destined to die upon exposure to taxol or okadaic acid. Specifically, TPA induces phosphorylation at a conserved extracellular signal-regulated kinase (ERK) site in the PEST region (Thr 163) and slows turnover of the normally rapidly degraded MCL1 protein; however, okadaic acid and taxol induce ERK-independent MCL1 phosphorylation at additional discrete sites. These findings add a new dimension to our understanding of the complex regulation of antiapoptotic BCL2 family members by demonstrating that, in addition to transcriptional and post-transcriptional regulation, MCL1 is subject to multiple, separate, post-translational phosphorylation events, produced in living versus dying cells at ERKinducible versus ERK-independent sites.

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Section: Resultsmentioning

confidence: 98%

Section: Cell Culture and Drug Treatmentmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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MCL1 is phosphorylated in the PEST region and stabilized upon ERK activation in viable cells, and at additional sites with cytotoxic okadaic acid or taxol

et al. 2004

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“…In contrast, the anti-apoptotic Bcl-2 family members are believed to function by dimerizing with pro-apoptotic forms and preventing cytochrome c release (Antonsson et al, 1997). Mcl-1 is an anti-apoptotic Bcl-2 family member that has been shown to prevent the onset of apoptosis in many cell types (Lomo et al, 1996;Reynolds et al, 1994;Zhou et al, 1997Zhou et al, , 1998). Numerous reports demonstrate that reduction in Mcl-1 protein levels using antisense is su cient to induce apoptosis (Chao et al, 1998;Leu et al, 2000;Moulding et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

Direct repression of the Mcl-1 promoter by E2F1

et al. 2002

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E2F1 induces apoptosis via both p53-dependent and p53-independent mechanisms. The direct targets in the p53-independent pathway remain enigmatic; however, the induction of this pathway does not require the transactivation domain of E2F1. Using cells that are defective in p53 activation, we show that E2F1 potently represses the expression of Mcl-1 ± an anti-apoptotic Bcl-2 family member whose depletion results in apoptosis. We also show that this transcriptional repression is direct and dependent upon E2F1's DNAbinding domain, but does not require the transactivation domain of E2F1. Consistent with this DNA binding requirement of E2F1 to repress Mcl-1, we show that E2F1 binds to the Mcl-1 promoter both in vitro and in vivo, and have identi®ed the DNA element (7143/ 7117) within this promoter that is required for E2F1 binding and repression. Additionally, cell lines constitutively expressing Mcl-1 are resistant to E2F1-mediated apoptosis ± suggesting that Mcl-1 downregulation is a necessary event in the p53-independent apoptotic process. Thus, we identify a p53 family-independent mechanism of E2F1-induced apoptosis in which E2F1 directly represses Mcl-1 expression.

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“…It contains two highly conserved regions, designated as the BH1 and BH2 domains, which have been shown to be required for the antiapoptotic function of Bcl-2 and for heterodimerization with Bax (Yin et al, 1994). Overexpression of Mcl-1 was found to delay apoptosis induced by various stimuli (Reynolds et al, 1994;Zhou et al, 1997;Leu et al, 2000). As a result of the similarity in sequence and function of Bcl-2 and Mcl-1, we have investigated whether or not PDT affects Mcl-1 expression and whether Pc 4-PDT can photodamage Mcl-1 as it does Bcl-2 and Bcl-xL.…”

mentioning

confidence: 99%