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Characterization of binding of LARP6 to the 5' stem-loop of collagen mRNAs

Title: Characterization of binding of LARP6 to the 5' stem-loop of collagen mRNAs: implications for synthesis of type I collagen.
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Name(s): Stefanovic, Lela, author
Longo, Liam, author
Zhang, Yujie, author
Stefanovic, Branko, author
Type of Resource: text
Genre: Journal Article
Text
Date Issued: 2014-01-01
Physical Form: computer
online resource
Extent: 1 online resource
Language(s): English
Abstract/Description: Type I collagen is composed of 2 polypeptides, α1(I) and α2(I), which fold into triple helix. Collagen α1(I) and α2(I) mRNAs have a conserved stem-loop structure in their 5' UTRs, the 5'SL. LARP6 binds the 5'SL to regulate type I collagen expression. We show that 5 nucleotides within the single stranded regions of 5'SL contribute to the high affinity of LARP6 binding. Mutation of individual nucleotides abolishes the binding in gel mobility shift assay. LARP6 binding to 5'SL of collagen α2(I) mRNA is more stable than the binding to 5'SL of α1(I) mRNA, although the equilibrium binding constants are similar. The more stable binding to α2(I) mRNA may favor synthesis of the heterotrimeric type I collagen. LARP6 needs 2 domains to contact 5'SL, the La domain and the RRM. T133 in the La domain is critical for folding of the protein, while loop 3 in the RRM is critical for binding 5'SL. Loop 3 is also involved in the interaction of LARP6 and protein translocation channel SEC61. This interaction is essential for type I collagen synthesis, because LARP6 mutant which binds 5'SL but which does not interact with SEC61, suppresses collagen synthesis in a dominant negative manner. We postulate that LARP6 directly targets collagen mRNAs to the SEC61 translocons to facilitate coordinated translation of the 2 collagen mRNAs. The unique sequences of LARP6 identified in this work may have evolved to enable its role in type I collagen biosynthesis.
Identifier: FSU_pmch_25692237 (IID), 10.1080/15476286.2014.996467 (DOI), PMC4615758 (PMCID), 25692237 (RID), 25692237 (EID)
Keywords: LARP6, RNA stem-loop, Fluorescence polarization, MRNA binding, Type I collagen
Grant Number: 2R01DK059466-07A2
Publication Note: This NIH-funded author manuscript originally appeared in PubMed Central at https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4615758.
Subject(s): 5' Untranslated Regions/genetics
Amino Acid Sequence
Autoantigens/genetics
Autoantigens/metabolism
Base Sequence
Binding Sites/genetics
Binding, Competitive
Collagen/genetics
Collagen/metabolism
Collagen Type I/genetics
Collagen Type I/metabolism
HEK293 Cells
Humans
Immunoblotting
Membrane Proteins/genetics
Membrane Proteins/metabolism
Molecular Sequence Data
Mutation
Nucleic Acid Conformation
Protein Binding
Protein Isoforms/genetics
Protein Isoforms/metabolism
Protein Stability
RNA, Messenger/chemistry
RNA, Messenger/genetics
RNA, Messenger/metabolism
RNA, Spliced Leader/chemistry
RNA, Spliced Leader/genetics
RNA, Spliced Leader/metabolism
Ribonucleoproteins/genetics
Ribonucleoproteins/metabolism
SEC Translocation Channels
Sequence Homology, Amino Acid
Persistent Link to This Record: http://purl.flvc.org/fsu/fd/FSU_pmch_25692237
Owner Institution: FSU
Is Part Of: RNA biology.
1555-8584
Issue: iss. 11, vol. 11

Choose the citation style.
Stefanovic, L., Longo, L., Zhang, Y., & Stefanovic, B. (2014). Characterization of binding of LARP6 to the 5' stem-loop of collagen mRNAs: implications for synthesis of type I collagen. Rna Biology. Retrieved from http://purl.flvc.org/fsu/fd/FSU_pmch_25692237