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 Hake, L. E.
Right arrow Articles by Richter, J. D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Hake, L. E.
Right arrow Articles by Richter, J. D.

 Previous Article  |  Next Article 

Mol Cell Biol, February 1998, p. 685-693, Vol. 18, No. 2
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Specificity of RNA Binding by CPEB: Requirement for RNA Recognition Motifs and a Novel Zinc Finger

Laura E. Hake,dagger Raul Mendez,Dagger and Joel D. Richter*

Worcester Foundation for Biomedical Research, Shrewsbury, Massachusetts 01545

Received 20 June 1997/Returned for modification 4 September 1997/Accepted 6 November 1997

CPEB is an RNA binding protein that interacts with the maturation-type cytoplasmic polyadenylation element (CPE) (consensus UUUUUAU) to promote polyadenylation and translational activation of maternal mRNAs in Xenopus laevis. CPEB, which is conserved from mammals to invertebrates, is composed of three regions: an amino-terminal portion with no obvious functional motif, two RNA recognition motifs (RRMs), and a cysteine-histidine region that is reminiscent of a zinc finger. In this study, we investigated the physical properties of CPEB required for RNA binding. CPEB can interact with RNA as a monomer, and phosphorylation, which modifies the protein during oocyte maturation, has little effect on RNA binding. Deletion mutations of CPEB have been overexpressed in Escherichia coli and used in a series of RNA gel shift experiments. Although a full-length and a truncated CPEB that lacks 139 amino-terminal amino acids bind CPE-containing RNA avidly, proteins that have had either RRM deleted bind RNA much less efficiently. CPEB that has had the cysteine-histidine region deleted has no detectable capacity to bind RNA. Single alanine substitutions of specific cysteine or histidine residues within this region also abolish RNA binding, pointing to the importance of this highly conserved domain of the protein. Chelation of metal ions by 1,10-phenanthroline inhibits the ability of CPEB to bind RNA; however, RNA binding is restored if the reaction is supplemented with zinc. CPEB also binds other metals such as cobalt and cadmium, but these destroy RNA binding. These data indicate that the RRMs and a zinc finger region of CPEB are essential for RNA binding.

* Corresponding author. Present address: Department of Molecular Genetics and Microbiology, University of Massachusetts Medical School, 222 Maple Ave., Shrewsbury, MA 01545. Phone: (508) 842-8921, ext. 340. Fax: (508) 842-3915. E-mail: joel.richter{at}banyan.ummed.edu.

dagger Present address: Department of Biology, Boston College, Chestnut Hill, MA 02167-3811.

Dagger Present address: Department of Molecular Genetics and Microbiology, University of Massachusetts Medical School, Shrewsbury, MA 01545.




This article has been cited by other articles:

  • Burns, D. M., Richter, J. D. (2008). CPEB regulation of human cellular senescence, energy metabolism, and p53 mRNA translation. Genes Dev. 22: 3449-3460 [Abstract] [Full Text]  
  • Guzeloglu-Kayisli, O., Pauli, S., Demir, H., Lalioti, M. D., Sakkas, D., Seli, E. (2008). Identification and characterization of human embryonic poly(A) binding protein (EPAB). Mol Hum Reprod 14: 581-588 [Abstract] [Full Text]  
  • Zearfoss, N. R., Alarcon, J. M., Trifilieff, P., Kandel, E., Richter, J. D. (2008). A Molecular Circuit Composed of CPEB-1 and c-Jun Controls Growth Hormone-Mediated Synaptic Plasticity in the Mouse Hippocampus. J. Neurosci. 28: 8502-8509 [Abstract] [Full Text]  
  • Keady, B. T., Kuo, P., Martinez, S. E., Yuan, L., Hake, L. E. (2007). MAPK interacts with XGef and is required for CPEB activation during meiosis in Xenopus oocytes. J. Cell Sci. 120: 1093-1103 [Abstract] [Full Text]  
  • Groisman, I., Ivshina, M., Marin, V., Kennedy, N. J., Davis, R. J., Richter, J. D. (2006). Control of cellular senescence by CPEB. Genes Dev. 20: 2701-2712 [Abstract] [Full Text]  
  • ROUHANA, L., WANG, L., BUTER, N., KWAK, J. E., SCHILTZ, C. A., GONZALEZ, T., KELLEY, A. E., LANDRY, C. F., WICKENS, M. (2005). Vertebrate GLD2 poly(A) polymerases in the germline and the brain. RNA 11: 1117-1130 [Abstract] [Full Text]  
  • Wilczynska, A., Aigueperse, C., Kress, M., Dautry, F., Weil, D. (2005). The translational regulator CPEB1 provides a link between dcp1 bodies and stress granules. J. Cell Sci. 118: 981-992 [Abstract] [Full Text]  
  • Alarcon, J. M., Hodgman, R., Theis, M., Huang, Y.-S., Kandel, E. R., Richter, J. D. (2004). Selective Modulation of Some Forms of Schaffer Collateral-CA1 Synaptic Plasticity in Mice With a Disruption of the CPEB-1 Gene. Learn. Mem. 11: 318-327 [Abstract] [Full Text]  
  • Minshall, N., Standart, N. (2004). The active form of Xp54 RNA helicase in translational repression is an RNA-mediated oligomer. Nucleic Acids Res 32: 1325-1334 [Abstract] [Full Text]  
  • Kurihara, Y., Tokuriki, M., Myojin, R., Hori, T., Kuroiwa, A., Matsuda, Y., Sakurai, T., Kimura, M., Hecht, N. B., Uesugi, S. (2003). CPEB2, A Novel Putative Translational Regulator in Mouse Haploid Germ Cells. Biol. Reprod. 69: 261-268 [Abstract] [Full Text]  
  • Huang, Y.-S., Carson, J. H., Barbarese, E., Richter, J. D. (2003). Facilitation of dendritic mRNA transport by CPEB. Genes Dev. 17: 638-653 [Abstract] [Full Text]  
  • Jin, S.-W., Arno, N., Cohen, A., Shah, A., Xu, Q., Chen, N., Ellis, R. E. (2001). In Caenorhabditis elegans, the RNA-Binding Domains of the Cytoplasmic Polyadenylation Element Binding Protein FOG-1 Are Needed to Regulate Germ Cell Fates. Genetics 159: 1617-1630 [Abstract] [Full Text]  
  • Dickson, K. S., Thompson, S. R., Gray, N. K., Wickens, M. (2001). Poly(A) Polymerase and the Regulation of Cytoplasmic Polyadenylation. J. Biol. Chem. 276: 41810-41816 [Abstract] [Full Text]  
  • Hodgman, R., Tay, J., Mendez, R., Richter, J. D. (2001). CPEB phosphorylation and cytoplasmic polyadenylation are catalyzed by the kinase IAK1/Eg2 in maturing mouse oocytes. Development 128: 2815-2822 [Abstract] [Full Text]  
  • Tan, L, Chang, J., Costa, A, Schedl, P (2001). An autoregulatory feedback loop directs the localized expression of the Drosophila CPEB protein Orb in the developing oocyte. Development 128: 1159-1169 [Abstract]  
  • WICKENS, M., BERNSTEIN, D., CRITTENDEN, S., LUITJENS, C., KIMBLE, J. (2001). PUF Proteins and 3'UTR Regulation in the Caenorhabditis elegans Germ Line. Cold Spring Harb Symp Quant Biol 66: 337-344 [Abstract]  
  • GROISMAN, I., HUANG, Y.-S., MENDEZ, R., CAO, Q., RICHTER, J.D. (2001). Translational Control of Embryonic Cell Division by CPEB and Maskin. Cold Spring Harb Symp Quant Biol 66: 345-352 [Abstract]  
  • Neto, O. P. d. M., Walker, J. A., Sa, C. M. d., Standart, N. (2000). Levels of free PABP are limited by newly polyadenylated mRNA in early Spisula embryogenesis. Nucleic Acids Res 28: 3346-3353 [Abstract] [Full Text]  
  • Richter, J. D. (1999). Cytoplasmic Polyadenylation in Development and Beyond. Microbiol. Mol. Biol. Rev. 63: 446-456 [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»