This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Mosialos, G Articles by Gilmore, T D Search for Related Content PubMed PubMed Citation Articles by Mosialos, G Articles by Gilmore, T D

A protein kinase-A recognition sequence is structurally linked to transformation by p59v-rel and cytoplasmic retention of p68c-rel.

Department of Chemistry, Boston University, Massachusetts 02215.

ABSTRACT

The Rel family of proteins includes a number of proteins involved in transcriptional control, such as the retroviral oncoprotein v-Rel, c-Rel, the Drosophila melanogaster developmental protein Dorsal, and subunits of the transcription factor NF-kappa B. These proteins are related through a highly conserved domain of approximately 300 amino acids, called the Rel homology domain, that contains dimerization, DNA binding, and nuclear targeting functions. Also within the Rel homology domain, there is a conserved consensus sequence (Arg-Arg-Pro-Ser) for phosphorylation by cyclic AMP-dependent protein kinase (PKA). We used linker insertion mutagenesis and site-directed mutagenesis to determine the importance of this sequence for the transformation of avian spleen cells by v-Rel and the subcellular localization of c-Rel in chicken embryo fibroblasts (CEF). The insertion of 2 amino acids (Pro-Trp) within this sequence completely abolished transformation and transcriptional repression by v-Rel and resulted in a shift in the localization of c-Rel from cytoplasmic to nuclear in CEF. When the conserved Ser within the PKA recognition sequence was replaced by Ala, there was no significant effect on transformation and transcriptional repression by v-Rel or on cytoplasmic retention of c-Rel. However, when this Ser was changed to Asp or Glu, transformation and transcriptional repression by v-Rel were significantly inhibited and c-Rel showed a diffuse nuclear and cytoplasmic localization in CEF. Although a peptide containing the recognition sequence from v-Rel can be phosphorylated by PKA in vitro, this site is not constitutively phosphorylated to a high degree in vivo in transformed spleen cells incubated with okadaic acid. Our results indicate that the transforming and transcriptional repressing activities of v-Rel and the cytoplasmic retention of c-Rel are dependent on the structure of the conserved PKA recognition motif. In addition, they suggest that phosphorylation at the conserved PKA site could have a negative effect on transformation and transcriptional repression by v-Rel and induce the nuclear localization of c-Rel.

This article has been cited by other articles:

• Johannessen, M., Delghandi, M. P., Rykx, A., Dragset, M., Vandenheede, J. R., Van Lint, J., Moens, U. (2007). Protein Kinase D Induces Transcription through Direct Phosphorylation of the cAMP-response Element-binding Protein. J. Biol. Chem. 282: 14777-14787 [Abstract] [Full Text]  
• Li, C., Wu, R.-C., Amazit, L., Tsai, S. Y., Tsai, M.-J., O'Malley, B. W. (2007). Specific Amino Acid Residues in the Basic Helix-Loop-Helix Domain of SRC-3 Are Essential for Its Nuclear Localization and Proteasome-Dependent Turnover. Mol. Cell. Biol. 27: 1296-1308 [Abstract] [Full Text]  
• Lu, K. T., Sinquett, F. L., Dryer, R. L., Song, C., Covey, L. R. (2006). c-Rel plays a key role in deficient activation of B cells from a non-X-linked hyper-IgM patient. Blood 108: 3769-3776 [Abstract] [Full Text]  
• Shin, I., Rotty, J., Wu, F. Y., Arteaga, C. L. (2005). Phosphorylation of p27Kip1 at Thr-157 Interferes with Its Association with Importin {alpha} during G1 and Prevents Nuclear Re-entry. J. Biol. Chem. 280: 6055-6063 [Abstract] [Full Text]  
• Rayet, B., Fan, Y., Gelinas, C. (2003). Mutations in the v-Rel Transactivation Domain Indicate Altered Phosphorylation and Identify a Subset of NF-{kappa}B-Regulated Cell Death Inhibitors Important for v-Rel Transforming Activity. Mol. Cell. Biol. 23: 1520-1533 [Abstract] [Full Text]  
• Antonsson, A., Hughes, K., Edin, S., Grundstrom, T. (2003). Regulation of c-Rel Nuclear Localization by Binding of Ca2+/Calmodulin. Mol. Cell. Biol. 23: 1418-1427 [Abstract] [Full Text]  
• Raval, A., Weissman, J. D., Howcroft, T. K., Singer, D. S. (2003). The GTP-Binding Domain of Class II Transactivator Regulates Its Nuclear Export. J. Immunol. 170: 922-930 [Abstract] [Full Text]  
• Liss, A. S., Bose, H. R. Jr. (2002). Mutational Analysis of the v-Rel Dimerization Interface Reveals a Critical Role for v-Rel Homodimers in Transformation. J. Virol. 76: 4928-4939 [Abstract] [Full Text]  
• Bose, H. R. Jr. (2001). Interferon Regulatory Factor 4 Contributes to Transformation of v-Rel-Expressing Fibroblasts. Mol. Cell. Biol. 21: 6369-6386 [Abstract] [Full Text]  
• Chen, C., Agnes, F., Gelinas, C. (1999). Mapping of a Serine-Rich Domain Essential for the Transcriptional, Antiapoptotic, and Transforming Activities of the v-Rel Oncoprotein. Mol. Cell. Biol. 19: 307-316 [Abstract] [Full Text]  
• Briggs, L. J., Stein, D., Goltz, J., Corrigan, V. C., Efthymiadis, A., Hubner, S., Jans, D. A. (1998). The cAMP-dependent Protein Kinase Site (Ser312) Enhances Dorsal Nuclear Import through Facilitating Nuclear Localization Sequence/Importin Interaction. J. Biol. Chem. 273: 22745-22752 [Abstract] [Full Text]  
• Wang, W., Rath, S., Durdik, J. M., Sen, R. (1998). Pentoxifylline Inhibits Ig {kappa} Gene Transcription and Rearrangements in Pre-B Cells. J. Immunol. 160: 1789-1795 [Abstract] [Full Text]  
• Hubner, S., Xiao, C.-Y., Jans, D. A. (1997). The Protein Kinase CK2 Site (Ser111/112) Enhances Recognition of the Simian Virus 40 Large T-antigen Nuclear Localization Sequence by Importin. J. Biol. Chem. 272: 17191-17195 [Abstract] [Full Text]  
• Xiao, C.-Y., Hübner, S., Elliot, R. M., Caon, A., Jans, D. A. (1996). A Consensus cAMP-dependent Protein Kinase (PK-A) Site in Place of the CcN Motif Casein Kinase II Site of Simian Virus 40 Large T-antigen Confers PK-A-mediated Regulation of Nuclear Import. J. Biol. Chem. 271: 6451-6457 [Abstract] [Full Text]  
• Verma, I M, Stevenson, J K, Schwarz, E M, Van Antwerp, D, Miyamoto, S (1995). Rel/NF-kappa B/I kappa B family: intimate tales of association and dissociation.. Genes Dev. 9: 2723-2735  
• Diehl, J. A., Tong, W., Sun, G., Hannink, M. (1995). Tumor Necrosis Factor-alpha-dependent Activation of a RelA Homodimer in Astrocytes. J. Biol. Chem. 270: 2703-2707 [Abstract] [Full Text]  
• Vandromme, M, Carnac, G, Gauthier-Rouviere, C, Fesquet, D, Lamb, N, Fernandez, A (1994). Nuclear import of the myogenic factor MyoD requires cAMP-dependent protein kinase activity but not the direct phosphorylation of MyoD. J. Cell Sci. 107: 613-620 [Abstract]  
• Norris, J L, Manley, J L (1992). Selective nuclear transport of the Drosophila morphogen dorsal can be established by a signaling pathway involving the transmembrane protein Toll and protein kinase A.. Genes Dev. 6: 1654-1667 [Abstract]