Identification and characterization of a novel RNA binding protein that associates with the 5′-untranslated region of the chloroplast psbA mRNA

Dwight Barnes, Amybeth Cohen, Richard K. Bruick, Katherine Kantardjieff, Sean Fowler, Ekem Efuet, Stephen P. Mayfield

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Binding of proteins to chloroplast-encoded mRNAs has been shown to be an essential part of chloroplast gene expression. Four nuclear-encoded proteins (38, 47, 55, and 60 kDa) have been identified that bind to the 5′-untranslated region of the Chlamydomonas reinhardtii psbA mRNA with high affinity and specificity. We have cloned a cDNA that represents the 38 kDa protein (RB38) and show that it encodes a novel RNA binding protein that is primarily localized within the chloroplast stroma. RB38 contains four 70 amino acid repeats with a high percentage of basic amino acids, as well as an amino-terminal extension predicted to act as a chloroplast import sequence. We demonstrate that the 38 kDa precursor protein is imported into isolated chloroplasts and interacts with high specificity to uridine-rich regions within the 5′-untranslated region of the psbA mRNA. While database searches have identified hypothetical proteins from several other eukaryotic species with high sequence similarity to the deduced amino acid sequence of RB38, no proteins with homology to RB38 have been biochemically characterized. Bioinformatic analysis of the RB38 sequence, together with structure analysis using circular dichroism and protein modeling, suggests that the 70 amino acid repeats within RB38 are similar in fold to previously identified RNA binding motifs, despite limited sequence homology.

Original languageEnglish (US)
Pages (from-to)8541-8550
Number of pages10
JournalBiochemistry
Volume43
Issue number26
DOIs
StatePublished - Jul 6 2004

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RNA-Binding Proteins
5' Untranslated Regions
Chloroplasts
Messenger RNA
Amino Acids
Proteins
Chloroplast Genes
Chlamydomonas reinhardtii
Basic Amino Acids
Protein Precursors
Uridine
Bioinformatics
Sequence Homology
Nuclear Proteins
Circular Dichroism
Computational Biology
Gene expression
Sequence Analysis
Amino Acid Sequence
Carrier Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Barnes, D., Cohen, A., Bruick, R. K., Kantardjieff, K., Fowler, S., Efuet, E., & Mayfield, S. P. (2004). Identification and characterization of a novel RNA binding protein that associates with the 5′-untranslated region of the chloroplast psbA mRNA. Biochemistry, 43(26), 8541-8550. https://doi.org/10.1021/bi035909j

Identification and characterization of a novel RNA binding protein that associates with the 5′-untranslated region of the chloroplast psbA mRNA. / Barnes, Dwight; Cohen, Amybeth; Bruick, Richard K.; Kantardjieff, Katherine; Fowler, Sean; Efuet, Ekem; Mayfield, Stephen P.

In: Biochemistry, Vol. 43, No. 26, 06.07.2004, p. 8541-8550.

Research output: Contribution to journalArticle

Barnes, D, Cohen, A, Bruick, RK, Kantardjieff, K, Fowler, S, Efuet, E & Mayfield, SP 2004, 'Identification and characterization of a novel RNA binding protein that associates with the 5′-untranslated region of the chloroplast psbA mRNA', Biochemistry, vol. 43, no. 26, pp. 8541-8550. https://doi.org/10.1021/bi035909j
Barnes, Dwight ; Cohen, Amybeth ; Bruick, Richard K. ; Kantardjieff, Katherine ; Fowler, Sean ; Efuet, Ekem ; Mayfield, Stephen P. / Identification and characterization of a novel RNA binding protein that associates with the 5′-untranslated region of the chloroplast psbA mRNA. In: Biochemistry. 2004 ; Vol. 43, No. 26. pp. 8541-8550.
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