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 Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Rubinfeld, B Articles by Polakis, P Search for Related Content PubMed PubMed Citation Articles by Rubinfeld, B Articles by Polakis, P

 Previous Article  |  Next Article 

Mol Cell Biol. 1992 October; 12(10): 4634-4642

Localization of the rap1GAP catalytic domain and sites of phosphorylation by mutational analysis.

Department of Molecular Biology, Onyx Pharmaceuticals, Emeryville, California.

ABSTRACT

rap1GAP is a GTPase-activating protein that specifically stimulates the GTP hydrolytic rate of p21rap1. We have defined the catalytic domain of rap1GAP by constructing a series of cDNAs coding for mutant proteins progressively deleted at the amino- and carboxy-terminal ends. Analysis of the purified mutant proteins shows that of 663 amino acid residues, only amino acids 75 to 416 are necessary for full GAP activity. Further truncation at the amino terminus resulted in complete loss of catalytic activity, whereas removal of additional carboxy-terminal residues dramatically accelerated the degradation of the protein in vivo. The catalytic domain we have defined excludes the region of rap1GAP which undergoes phosphorylation on serine residues. We have further defined this phosphoacceptor region of rap1GAP by introducing point mutations at specific serine residues and comparing the phosphopeptide maps of the mutant proteins. Two of the sites of phosphorylation by cyclic AMP (cAMP)-dependent kinase were localized to serine residues 490 and 499, and one site of phosphorylation by p34cdc2 was localized to serine 484. In vivo, rap1GAP undergoes phosphorylation at four distinct sites, two of which appear to be identical to the sites phosphorylated by cAMP-dependent kinase in vitro.

This article has been cited by other articles:

• Schultess, J., Danielewski, O., Smolenski, A. P. (2005). Rap1GAP2 is a new GTPase-activating protein of Rap1 expressed in human platelets. Blood 105: 3185-3192 [Abstract] [Full Text]  
• Krymskaya, V. P., Shipley, J. M. (2003). Lymphangioleiomyomatosis: A Complex Tale of Serum Response Factor-Mediated Tissue Inhibitor of Metalloproteinase-3 Regulation. Am. J. Respir. Cell Mol. Bio. 28: 546-550 [Full Text]  
• Norum, J. H., Hart, K., Levy, F. O. (2003). Ras-dependent ERK Activation by the Human Gs-coupled Serotonin Receptors 5-HT4(b) and 5-HT7(a). J. Biol. Chem. 278: 3098-3104 [Abstract] [Full Text]  
• Singh, L., Gao, Q., Kumar, A., Gotoh, T., Wazer, D. E., Band, H., Feig, L. A., Band, V. (2002). The High-Risk Human Papillomavirus Type 16 E6 Counters the GAP Function of E6TP1 toward Small Rap G Proteins. J. Virol. 77: 1614-1620 [Abstract] [Full Text]  
• Meng, J., Casey, P. J. (2002). Activation of Gz Attenuates Rap1-mediated Differentiation of PC12 Cells. J. Biol. Chem. 277: 43417-43424 [Abstract] [Full Text]  
• Brinkmann, T., Daumke, O., Herbrand, U., Kuhlmann, D., Stege, P., Ahmadian, M. R., Wittinghofer, A. (2002). Rap-specific GTPase Activating Protein follows an Alternative Mechanism. J. Biol. Chem. 277: 12525-12531 [Abstract] [Full Text]  
• Meng, J., Glick, J. L., Polakis, P., Casey, P. J. (1999). Functional Interaction between Galpha z and Rap1GAP Suggests a Novel Form of Cellular Cross-talk. J. Biol. Chem. 274: 36663-36669 [Abstract] [Full Text]  
• Gao, Q., Srinivasan, S., Boyer, S. N., Wazer, D. E., Band, V. (1999). The E6 Oncoproteins of High-Risk Papillomaviruses Bind to a Novel Putative GAP Protein, E6TP1, and Target It for Degradation. Mol. Cell. Biol. 19: 733-744 [Abstract] [Full Text]  
• Guasch, R. M., Scambler, P., Jones, G. E., Ridley, A. J. (1998). RhoE Regulates Actin Cytoskeleton Organization and Cell Migration. Mol. Cell. Biol. 18: 4761-4771 [Abstract] [Full Text]  
• Hu, Y., Benedict, M. A., Wu, D., Inohara, N., Nunez, G. (1998). Bcl-XL interacts with Apaf-1 and inhibits Apaf-1-dependent caspase-9 activation. Proc. Natl. Acad. Sci. USA 95: 4386-4391 [Abstract] [Full Text]  
• Chen, F., Barkett, M., Ram, K. T., Quintanilla, A., Hariharan, I. K. (1997). Biological characterization of Drosophila Rapgap1, a GTPase activating protein for Rap1. Proc. Natl. Acad. Sci. USA 94: 12485-12490 [Abstract] [Full Text]  
• Kurachi, H., Wada, Y., Tsukamoto, N., Maeda, M., Kubota, H., Hattori, M., Iwai, K., Minato, N. (1997). Human SPA-1 Gene Product Selectively Expressed in Lymphoid Tissues Is a Specific GTPase-activating Protein for Rap1 and Rap2. SEGREGATE EXPRESSION PROFILES FROM A rap1GAP GENE PRODUCT. J. Biol. Chem. 272: 28081-28088 [Abstract] [Full Text]  
• Sun, X. J., Pons, S., Asano, T., Myers, M. G. Jr., Glasheen, E., White, M. F. (1996). The Fyn Tyrosine Kinase Binds Irs-1 and Forms a Distinct Signaling Complex during Insulin Stimulation. J. Biol. Chem. 271: 10583-10587 [Abstract] [Full Text]  
• Wienecke, R., König, A., DeClue, J. E. (1995). Identification of Tuberin, the Tuberous Sclerosis-2 Product. J. Biol. Chem. 270: 16409-16414 [Abstract] [Full Text]  
• Altschuler, D. L., Peterson, S. N., Ostrowski, M. C., Lapetina, E. G. (1995). Cyclic AMP-dependent Activation of Rap1b. J. Biol. Chem. 270: 10373-10376 [Abstract] [Full Text]  
• Rubinfeld, B., Souza, B., Albert, I., Munemitsu, S., Polakis, P. (1995). The APC Protein and E-cadherin Form Similar but Independent Complexes with alpha-Catenin, beta-Catenin, and Plakoglobin. J. Biol. Chem. 270: 5549-5555 [Abstract] [Full Text]  
• Zhu, J., Reynet, C., Caldwell, J. S., Kahn, C. R. (1995). Characterization of Rad, a New Member of Ras/GTPase Superfamily, and Its Regulation by a Unique GTPase-activating protein (GAP)-like Activity. J. Biol. Chem. 270: 4805-4812 [Abstract] [Full Text]  
• Rubinfeld, B, Souza, B, Albert, I, Muller, O, Chamberlain, S., Masiarz, F., Munemitsu, S, Polakis, P (1993). Association of the APC gene product with beta-catenin. Science 262: 1731-1734 [Abstract]