Advertisement
JBC

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Originally published In Press as doi:10.1074/jbc.M102891200 on May 7, 2001

J. Biol. Chem., Vol. 276, Issue 29, 27188-27196, July 20, 2001
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
276/29/27188    most recent
M102891200v1
Right arrow Submit a Letter to Editor
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
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 arrowRequest Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Ohndorf, U.-M.
Right arrow Articles by Sondermann, P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ohndorf, U.-M.
Right arrow Articles by Sondermann, P.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

Contributions of the Individual Domains in Human La Protein to Its RNA 3'-End Binding Activity*

Uta-Maria OhndorfDagger , Clemens Steegborn, Rainer Knijff, and Peter Sondermann

From the Max-Planck-Institut für Biochemie, Abteilung Strukturforschung, Am Klopferspitz 18a, D-82152 Planegg-Martinsried, Germany

The autoantigen La regulates the maturation of RNA polymerase III transcripts by binding to their poly(U) termination signal. The modular protein harbors a N-terminal RNA recognition motif (RRM), RRM1, and in the C-terminal domain, a second, atypical RRM2, in addition to a phosphorylation site, and a putative nucleotide binding site. This study presents a detailed investigation into the RNA 3'-end binding properties of La by using binding titration and competition assays with subsequent gel mobility shift analysis. Two truncation mutants containing one (La-RRM1) or both (La-RRM1-RRM2) RNA-binding domains were constructed, overexpressed, and purified. A Kd value of 25 ± 10 nM for La binding to a nonameric RNA ligand with the oligouridylate recognition sequence was obtained, discriminating with a specificity ratio of ~100 for this probe over a RNA ligand with a 3'-poly(A) stretch. The N-terminal La-RRM1 region was identified as the major contributor of these properties to La, manifested in a 5-fold lower Kd of 5 ± 3 nM and a slightly increased specificity ratio of 120 for the RNA ligand. The atypical RRM2 in the C-terminal domain of La has an unprecedented negative effect on 3'-end RNA recognition, as indicated by a higher Kd value of 90 ± 10 nM for the La-RRM1-RRM2 mutant but comparable specificity. Thus the C-terminal regions beyond RRM2 positively modulate the RNA binding affinity of La. Negative regulation, however, occurs through Ser366 phosphorylation decreasing the binding affinity by 2-fold. ATP had no influence on RNA complex formation. The functional implications of these findings for the mechanism of action of La are discussed.

* The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger To whom correspondence should be addressed: Max-Planck-Institut für Biochemie, Abteilung Strukturforschung, Am Klopferspitz 18a, D-82152 Planegg-Martinsried, Germany. Fax: 49-89-8578-3516; E-mail: ohndorf@biochem.mpg.de.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 J. Virol.Home page
T. Mondal, U. Ray, A. K. Manna, R. Gupta, S. Roy, and S. Das
Structural Determinant of Human La Protein Critical for Internal Initiation of Translation of Hepatitis C Virus RNA
J. Virol., December 1, 2008; 82(23): 11927 - 11938.
[Abstract] [Full Text] [PDF]

Home page
 RNAHome page
K. Nykamp, M.-H. Lee, and J. Kimble
C. elegans La-related protein, LARP-1, localizes to germline P bodies and attenuates Ras-MAPK signaling during oogenesis
RNA, July 1, 2008; 14(7): 1378 - 1389.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
C. M. O'Connor and K. Collins
A Novel RNA Binding Domain in Tetrahymena Telomerase p65 Initiates Hierarchical Assembly of Telomerase Holoenzyme.
Mol. Cell. Biol., March 1, 2006; 26(6): 2029 - 2036.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
L. Huang, J. Hwang, S. D. Sharma, M. R. S. Hargittai, Y. Chen, J. J. Arnold, K. D. Raney, and C. E. Cameron
Hepatitis C Virus Nonstructural Protein 5A (NS5A) Is an RNA-binding Protein
J. Biol. Chem., October 28, 2005; 280(43): 36417 - 36428.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
F. Brenet, N. Dussault, J. Borch, G. Ferracci, C. Delfino, P. Roepstorff, R. Miquelis, and L. Ouafik
Mammalian Peptidylglycine {alpha}-Amidating Monooxygenase mRNA Expression Can Be Modulated by the La Autoantigen
Mol. Cell. Biol., September 1, 2005; 25(17): 7505 - 7521.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
R. V. Intine, M. Dundr, A. Vassilev, E. Schwartz, Y. Zhao, Y. Zhao, M. L. DePamphilis, and R. J. Maraia
Nonphosphorylated Human La Antigen Interacts with Nucleolin at Nucleolar Sites Involved in rRNA Biogenesis
Mol. Cell. Biol., December 15, 2004; 24(24): 10894 - 10904.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
S. Horke, K. Reumann, C. Schulze, F. Grosse, and T. Heise
The La Motif and the RNA Recognition Motifs of Human La Autoantigen Contribute Individually to RNA Recognition and Subcellular Localization
J. Biol. Chem., November 26, 2004; 279(48): 50302 - 50309.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
E. I. Schwartz, R. V. Intine, and R. J. Maraia
CK2 Is Responsible for Phosphorylation of Human La Protein Serine-366 and Can Modulate rpL37 5'-Terminal Oligopyrimidine mRNA Metabolism
Mol. Cell. Biol., November 1, 2004; 24(21): 9580 - 9591.
[Abstract] [Full Text] [PDF]

Home page
 GENES CELLSHome page
N. Hino, T. Suzuki, T. Yasukawa, K. Seio, K. Watanabe, and T. Ueda
The pathogenic A4269G mutation in human mitochondrial tRNAIle alters the T-stem structure and decreases the binding affinity for elongation factor Tu
Genes Cells, March 1, 2004; 9(3): 243 - 252.
[Abstract] [Full Text] [PDF]

Home page
 Eukaryot CellHome page
K. Tanabe, N. Ito, T. Wakuri, F. Ozoe, M. Umeda, S. Katayama, K. Tanaka, H. Matsuda, and M. Kawamukai
Sla1, a Schizosaccharomyces pombe Homolog of the Human La Protein, Induces Ectopic Meiosis when Its C Terminus Is Truncated
Eukaryot. Cell, December 1, 2003; 2(6): 1274 - 1287.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
R. Pudi, S. Abhiman, N. Srinivasan, and S. Das
Hepatitis C Virus Internal Ribosome Entry Site-mediated Translation Is Stimulated by Specific Interaction of Independent Regions of Human La Autoantigen
J. Biol. Chem., March 28, 2003; 278(14): 12231 - 12240.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
S. Horke, K. Reumann, A. Rang, and T. Heise
Molecular Characterization of the Human La Protein{middle dot}Hepatitis B Virus RNA.B Interaction in Vitro
J. Biol. Chem., September 13, 2002; 277(38): 34949 - 34958.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
 All ASBMB Journals   Molecular and Cellular Proteomics 
 Journal of Lipid Research   ASBMB Today 
Copyright © 2001 by the American Society for Biochemistry and Molecular Biology.
Advertisement
spacer
Advertisement
Advertisement