ScienceDirect - Cell : Intracellular Nucleotides Act = as Critical Prosurvival Factors by Binding to Cytochrome C and = Inhibiting Apoptosome

Intracellular Nucleotides Act as Critical Prosurvival Factors by = Binding to=20 Cytochrome C and Inhibiting Apoptosome

Dhyan Chandra¹^,=20^,, Shawn B. Bratton²,=20 Maria D. Person²,=20 Yanan Tian³,=20 Angel G. Martin⁴,=20 Mary Ayres⁵,=20 Howard O. Fearnhead⁴,=20 Varsha Gandhi⁵=20 and Dean G. Tang¹^,=20⁶^,=20^,

¹Department of=20 Carcinogenesis, University of Texas MD Anderson Cancer Center, Science=20 Park-Research Division, Smithville, TX 78957, USA
²Division of Pharmacology/Toxicology, College = of=20 Pharmacy, University of Texas at Austin, Austin, TX 78712, USA
³Department of Veterinary Physiology and = Pharmacology,=20 MS 4466, Texas A&M University, College Station, TX 77843, USA
⁴Apoptosis Section, Laboratory of Protein = Dynamics and=20 Signaling, NCI-Frederick, Frederick, MD 21702, USA
⁵Department of Experimental Therapeutics, = University of=20 Texas M. D. Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, TX = 77030,=20 USA
⁶Program in Molecular = Carcinogenesis of=20 Graduate School for Biomedical Science, University of Texas M. D. = Anderson=20 Cancer Center, 1515 Holcombe Boulevard, Houston, TX 77030, USA =

Received=20 22 June 2005; revised 6 January 2006; accepted 3 May = 2006. =20 Published: June 29, 2006. Available online 29 June 2006.

Summary

Cytochrome c (CC)-initiated Apaf-1 apoptosome formation represents a = key=20 initiating event in apoptosis. This process can be reconstituted in = vitro with=20 the addition of CC and ATP or dATP to cell lysates. How physiological = levels of=20 nucleotides, normally at high mM concentrations, affect apoptosome = activation=20 remains unclear. Here we show that physiological levels of nucleotides = inhibit=20 the CC-initiated apoptosome formation and caspase-9 activation by = directly=20 binding to CC on several key lysine residues and thus preventing CC = interaction=20 with Apaf-1. We show that in various apoptotic systems caspase = activation is=20 preceded or accompanied by decreases in overall intracellular NTP pools. = Microinjection of nucleotides inhibits whereas experimentally reducing = NTP pools=20 enhances both CC and apoptotic stimuli-induced cell death. Our results = thus=20 suggest that the intracellular nucleotides represent critical = prosurvival=20 factors by functioning as natural inhibitors of apoptosome formation and = a=20 barrier that cells must overcome the nucleotide barrier to undergo = apoptosis=20 cell death.
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Cell=20
Volume=20 125, Issue 7 , 30 June 2006, Pages 1333-1346=20

Copyright =C2=A9 = 2006 Elsevier = B.V. All rights=20 reserved. ScienceDirect=C2=AE is a registered trademark of = Elsevier=20 B.V.

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