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Defining the roles and interactions of PTB.
Biochem Soc Trans. 2012 Aug; 40(4):815-20.BS

Abstract

PTB (polypyrimidine tract-binding protein) is an abundant and widely expressed RNA-binding protein with four RRM (RNA recognition motif) domains. PTB is involved in numerous post-transcriptional steps in gene expression in both the nucleus and cytoplasm, but has been best characterized as a regulatory repressor of some ASEs (alternative splicing events), and as an activator of translation driven by IRESs (internal ribosome entry segments). We have used a variety of approaches to characterize the activities of PTB and its molecular interactions with RNA substrates and protein partners. Using splice-sensitive microarrays we found that PTB acts not only as a splicing repressor but also as an activator, and that these two activities are determined by the location at which PTB binds relative to target exons. We have identified minimal splicing repressor and activator domains, and have determined high resolution structures of the second RRM domain of PTB binding to peptide motifs from the co-repressor protein Raver1. Using single-molecule techniques we have determined the stoichiometry of PTB binding to a regulated splicing substrate in whole nuclear extracts. Finally, we have used tethered hydroxyl radical probing to determine the locations on viral IRESs at which each of the four RRM domains bind. We are now combining tethered probing with single molecule analyses to gain a detailed understanding of how PTB interacts with pre-mRNA substrates to effect either repression or activation of splicing.

Authors+Show Affiliations

Department of Biochemistry, University of Cambridge, Cambridge, UK.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Review

Language

eng

PubMed ID

22817740

Citation

Kafasla, Panagiota, et al. "Defining the Roles and Interactions of PTB." Biochemical Society Transactions, vol. 40, no. 4, 2012, pp. 815-20.
Kafasla P, Mickleburgh I, Llorian M, et al. Defining the roles and interactions of PTB. Biochem Soc Trans. 2012;40(4):815-20.
Kafasla, P., Mickleburgh, I., Llorian, M., Coelho, M., Gooding, C., Cherny, D., Joshi, A., Kotik-Kogan, O., Curry, S., Eperon, I. C., Jackson, R. J., & Smith, C. W. (2012). Defining the roles and interactions of PTB. Biochemical Society Transactions, 40(4), 815-20. https://doi.org/10.1042/BST20120044
Kafasla P, et al. Defining the Roles and Interactions of PTB. Biochem Soc Trans. 2012;40(4):815-20. PubMed PMID: 22817740.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Defining the roles and interactions of PTB. AU - Kafasla,Panagiota, AU - Mickleburgh,Ian, AU - Llorian,Miriam, AU - Coelho,Miguel, AU - Gooding,Clare, AU - Cherny,Dmitry, AU - Joshi,Amar, AU - Kotik-Kogan,Olga, AU - Curry,Stephen, AU - Eperon,Ian C, AU - Jackson,Richard J, AU - Smith,Christopher W J, PY - 2012/7/24/entrez PY - 2012/7/24/pubmed PY - 2012/12/12/medline SP - 815 EP - 20 JF - Biochemical Society transactions JO - Biochem. Soc. Trans. VL - 40 IS - 4 N2 - PTB (polypyrimidine tract-binding protein) is an abundant and widely expressed RNA-binding protein with four RRM (RNA recognition motif) domains. PTB is involved in numerous post-transcriptional steps in gene expression in both the nucleus and cytoplasm, but has been best characterized as a regulatory repressor of some ASEs (alternative splicing events), and as an activator of translation driven by IRESs (internal ribosome entry segments). We have used a variety of approaches to characterize the activities of PTB and its molecular interactions with RNA substrates and protein partners. Using splice-sensitive microarrays we found that PTB acts not only as a splicing repressor but also as an activator, and that these two activities are determined by the location at which PTB binds relative to target exons. We have identified minimal splicing repressor and activator domains, and have determined high resolution structures of the second RRM domain of PTB binding to peptide motifs from the co-repressor protein Raver1. Using single-molecule techniques we have determined the stoichiometry of PTB binding to a regulated splicing substrate in whole nuclear extracts. Finally, we have used tethered hydroxyl radical probing to determine the locations on viral IRESs at which each of the four RRM domains bind. We are now combining tethered probing with single molecule analyses to gain a detailed understanding of how PTB interacts with pre-mRNA substrates to effect either repression or activation of splicing. SN - 1470-8752 UR - https://www.unboundmedicine.com/medline/citation/22817740/Defining_the_roles_and_interactions_of_PTB_ L2 - https://portlandpress.com/biochemsoctrans/article-lookup/doi/10.1042/BST20120044 DB - PRIME DP - Unbound Medicine ER -