This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Peters, G. A.
Right arrow Articles by Sen, G. C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Peters, G. A.
Right arrow Articles by Sen, G. C.

 Previous Article  |  Next Article 

Molecular and Cellular Biology, March 2001, p. 1908-1920, Vol. 21, No. 6
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.6.1908-1920.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Modular Structure of PACT: Distinct Domains for Binding and Activating PKR

Gregory A. Peters,1,2 Rune Hartmann,1,dagger Jun Qin,3 and Ganes C. Sen1,2,*

Department of Molecular Biology1 and Department of Molecular Cardiology,3 Lerner Research Institute, The Cleveland Clinic Foundation, Cleveland, Ohio 44195, and Department of Physiology and Biophysics, School of Medicine, Case Western Reserve University, Cleveland, Ohio 441062

Received 3 August 2000/Returned for modification 11 September 2000/Accepted 6 December 2000

PACT is a 35-kDa human protein that can directly bind and activate the latent protein kinase, PKR. Here we report that PKR activation by PACT causes cellular apoptosis in addition to PKR autophosphorylation and translation inhibition. We analyzed the structure-function relationship of PACT by measuring its ability to bind and activate PKR in vitro and in vivo. Our studies revealed that among three domains of PACT, the presence of either domain 1 or domain 2 was sufficient for high-affinity binding of PACT to PKR. On the other hand, domain 3, consisting of 66 residues, was absolutely required for PKR activation in vitro and in vivo. When fused to maltose-binding protein, domain 3 was also sufficient for efficiently activating PKR in vitro. However, it bound poorly to PKR at the physiological salt concentration and consequently could not activate it properly in vivo. As anticipated, activation of PKR by domain 3 in vivo could be restored by attaching it to a heterologous PKR-binding domain. These results demonstrated that the structure of PACT is modular: it is composed of a distinct PKR-activation domain and two mutually redundant PKR-interacting domains.

* Corresponding author. Mailing address: Department of Molecular Biology/NC20, The Cleveland Clinic Foundation, 9500 Euclid Ave., Cleveland, OH 44195. Phone: (216) 444-0636. Fax: (216) 444-0513. E-mail: seng{at}ccf.org.

dagger Present address: Aarhus University, Aarhus, Denmark.

Molecular and Cellular Biology, March 2001, p. 1908-1920, Vol. 21, No. 6
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.6.1908-1920.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Peters, G. A., Seachrist, D. D., Keri, R. A., Sen, G. C. (2009). The double-stranded RNA-binding protein, PACT, is required for postnatal anterior pituitary proliferation. Proc. Natl. Acad. Sci. USA 106: 10696-10701 [Abstract] [Full Text]  
  • Daher, A., Laraki, G., Singh, M., Melendez-Pena, C. E., Bannwarth, S., Peters, A. H. F. M., Meurs, E. F., Braun, R. E., Patel, R. C., Gatignol, A. (2009). TRBP Control of PACT-Induced Phosphorylation of Protein Kinase R Is Reversed by Stress. Mol. Cell. Biol. 29: 254-265 [Abstract] [Full Text]  
  • Armstrong, M. E., Gantier, M., Li, L., Chung, W. Y., McCann, A., Baugh, J. A., Donnelly, S. C. (2008). Small Interfering RNAs Induce Macrophage Migration Inhibitory Factor Production and Proliferation in Breast Cancer Cells via a Double-Stranded RNA-Dependent Protein Kinase-Dependent Mechanism. J. Immunol. 180: 7125-7133 [Abstract] [Full Text]  
  • Mittelstadt, M., Frump, A., Khuu, T., Fowlkes, V., Handy, I., Patel, C. V., Patel, R. C. (2008). Interaction of human tRNA-dihydrouridine synthase-2 with interferon-induced protein kinase PKR. Nucleic Acids Res 36: 998-1008 [Abstract] [Full Text]  
  • Alirezaei, M., Watry, D. D., Flynn, C. F., Kiosses, W. B., Masliah, E., Williams, B. R. G., Kaul, M., Lipton, S. A., Fox, H. S. (2007). Human Immunodeficiency Virus-1/Surface Glycoprotein 120 Induces Apoptosis through RNA-Activated Protein Kinase Signaling in Neurons. J. Neurosci. 27: 11047-11055 [Abstract] [Full Text]  
  • Kok, K. H., Ng, M.-H. J., Ching, Y.-P., Jin, D.-Y. (2007). Human TRBP and PACT Directly Interact with Each Other and Associate with Dicer to Facilitate the Production of Small Interfering RNA. J. Biol. Chem. 282: 17649-17657 [Abstract] [Full Text]  
  • Yang, M., Wu, S., Su, X., May, W. S. (2006). JAZ mediates G1 cell-cycle arrest and apoptosis by positively regulating p53 transcriptional activity. Blood 108: 4136-4145 [Abstract] [Full Text]  
  • Garcia, M. A., Gil, J., Ventoso, I., Guerra, S., Domingo, E., Rivas, C., Esteban, M. (2006). Impact of Protein Kinase PKR in Cell Biology: from Antiviral to Antiproliferative Action. Microbiol. Mol. Biol. Rev. 70: 1032-1060 [Abstract] [Full Text]  
  • Peters, G. A., Li, S., Sen, G. C. (2006). Phosphorylation of Specific Serine Residues in the PKR Activation Domain of PACT Is Essential for Its Ability to Mediate Apoptosis. J. Biol. Chem. 281: 35129-35136 [Abstract] [Full Text]  
  • Bennett, R. L., Blalock, W. L., Abtahi, D. M., Pan, Y., Moyer, S. A., May, W. S. (2006). RAX, the PKR activator, sensitizes cells to inflammatory cytokines, serum withdrawal, chemotherapy, and viral infection. Blood 108: 821-829 [Abstract] [Full Text]  
  • Li, S., Peters, G. A., Ding, K., Zhang, X., Qin, J., Sen, G. C. (2006). Molecular basis for PKR activation by PACT or dsRNA. Proc. Natl. Acad. Sci. USA 103: 10005-10010 [Abstract] [Full Text]  
  • Chen, G., Ma, C., Bower, K. A., Ke, Z., Luo, J. (2006). Interaction between RAX and PKR Modulates the Effect of Ethanol on Protein Synthesis and Survival of Neurons. J. Biol. Chem. 281: 15909-15915 [Abstract] [Full Text]  
  • Parker, G. S., Eckert, D. M., Bass, B. L. (2006). RDE-4 preferentially binds long dsRNA and its dimerization is necessary for cleavage of dsRNA to siRNA. RNA 12: 807-818 [Abstract] [Full Text]  
  • Rowe, T. M., Rizzi, M., Hirose, K., Peters, G. A., Sen, G. C. (2006). A role of the double-stranded RNA-binding protein PACT in mouse ear development and hearing. Proc. Natl. Acad. Sci. USA 103: 5823-5828 [Abstract] [Full Text]  
  • Bennett, R. L., Blalock, W. L., May, W. S. (2004). Serine 18 Phosphorylation of RAX, the PKR Activator, Is Required for PKR Activation and Consequent Translation Inhibition. J. Biol. Chem. 279: 42687-42693 [Abstract] [Full Text]  
  • Buck, E., Iyengar, R. (2003). Organization and Functions of Interacting Domains for Signaling by Protein-Protein Interactions. Sci Signal 2003: re14-re14 [Abstract] [Full Text]  
  • Yang, M., Ito, T., May, W. S. (2003). A Novel Role for RAX, the Cellular Activator of PKR, in Synergistically Stimulating SV40 Large T Antigen-dependent Gene Expression. J. Biol. Chem. 278: 38325-38332 [Abstract] [Full Text]  
  • SAUNDERS, L. R., BARBER, G. N. (2003). The dsRNA binding protein family: critical roles, diverse cellular functions. FASEB J. 17: 961-983 [Abstract] [Full Text]  
  • Peters, G. A., Khoo, D., Mohr, I., Sen, G. C. (2002). Inhibition of PACT-Mediated Activation of PKR by the Herpes Simplex Virus Type 1 Us11 Protein. J. Virol. 76: 11054-11064 [Abstract] [Full Text]  
  • Gupta, V., Patel, R. C. (2002). Proapoptotic protein PACT is expressed at high levels in colonic epithelial cells in mice. Am. J. Physiol. Gastrointest. Liver Physiol. 283: G801-G808 [Abstract] [Full Text]  
  • Parker, L. M., Fierro-Monti, I., Mathews, M. B. (2001). Nuclear Factor 90 Is a Substrate and Regulator of the Eukaryotic Initiation Factor 2 Kinase Double-stranded RNA-activated Protein Kinase. J. Biol. Chem. 276: 32522-32530 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»