2MPU

Structural and Functional analysis of the Hordeum vulgare L. HvGR-RBP1 protein, a glycine-rich RNA binding protein implicated in the regulation of barley leaf senescence and environmental adaptation

Summary for 2MPU

• Entry DOI: 10.2210/pdb2mpu/pdb
• NMR Information: BMRB: 18776
• Descriptor: RBP1 (1 entity in total)
• Functional Keywords: rna recognition motif, rrm, rnp1, rnp2, glycine rich protein, nucleic acid binding protein, rna binding protein
• Biological source: Hordeum vulgare (barley)
• Total number of polymer chains: 1
• Total formula weight: 9989.83

Authors

Mason, K.E.,Tripet, B.P.,Eilers, B.J.,Powell, P.,Fischer, A.M.,Copie, V. (deposition date: 2014-06-02, release date: 2014-12-17, Last modification date: 2024-05-15)

Primary citation

Tripet, B.P.,Mason, K.E.,Eilers, B.J.,Burns, J.,Powell, P.,Fischer, A.M.,Copie, V.
Structural and Biochemical Analysis of the Hordeum vulgare L. HvGR-RBP1 Protein, a Glycine-Rich RNA-Binding Protein Involved in the Regulation of Barley Plant Development and Stress Response.
Biochemistry, 53:7945-7960, 2014

PubMed Abstract: The timing of whole-plant senescence influences important agricultural traits such as yield and grain protein content. Post-transcriptional regulation by plant RNA-binding proteins is essential for proper control of gene expression, development, and stress responses. Here, we report the three-dimensional solution NMR structure and nucleic acid-binding properties of the barley glycine-rich RNA-binding protein HvGR-RBP1, whose transcript has been identified as being >45-fold up-regulated in early-as compared to late-senescing near-isogenic barley germplasm. NMR analysis reveals that HvGR-RBP1 is a multidomain protein comprising a well-folded N-terminal RNA Recognition Motif (RRM) and a structurally disordered C-terminal glycine-rich domain. Chemical shift differences observed in 2D (1)H-(15)N correlation (HSQC) NMR spectra of full-length HvGR-RBP1 and N-HvGR-RBP1 (RRM domain only) suggest that the two domains can interact both in-trans and intramolecularly, similar to what is observed in the tobacco NtGR-RBP1 protein. Further, we show that the RRM domain of HvGR-RBP1 binds single-stranded DNA nucleotide fragments containing the consensus nucleotide sequence 5'-TTCTGX-3' with low micromolar affinity in vitro. We also demonstrate that the C-terminal glycine-rich (HvGR) domain of Hv-GR-RBP1 can interact nonspecifically with ssRNA in vitro. Structural similarities with other plant glycine-rich RNA-binding proteins suggest that HvGR-RBP1 may be multifunctional. Based on gene expression analysis following cold stress in barley and E. coli growth studies following cold shock treatment, we conclude that HvGR-RBP1 functions in a manner similar to cold-shock proteins and harbors RNA chaperone activity. HvGR-RBP1 is therefore not only involved in the regulation of barley development including senescence, but also functions in plant responses to environmental stress.

PubMed: 25495582
DOI: 10.1021/bi5007223

Experimental method

SOLUTION NMR