The 1H, 15N and 13C resonance assignments of the C-terminal domain of Serpine mRNA binding protein 1 (SERBP1)

Baudin, Antoine; Xu, Xiaoping; Libich, David S.

doi:10.1007/s12104-021-10046-3

Cited by 2 publications

(2 citation statements)

References 25 publications

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“…The SERBP1 189-400 construct (Addgene accession number 172315) was expressed in E.coli as previously described (Baudin et al, 2021). The genes for full-length SERBP1 and a SERBP1 149-400 construct were amplified by PCR (primer are shown in Supplementary Table S1), digested by KasI and BamHI restriction enzymes and cloned into a custom pAG8Ha-His vector that introduced an 8x histidine tag followed by a TEV cleavage site N-terminal to the coding sequence.…”

Section: Protein Expression and Purificationmentioning

confidence: 99%

“…Expression cultures were grown at 37 °C in baffled Fernbach flasks with shaking, and protein expression was induced at OD 600 ∼0.6-0.8 with 1 mM IPTG and continued for 3 or 6 h, for LB or M9 cultures, respectively. The cells were harvested by centrifugation at 4000 g for 20 min and the resulting pellets were stored at −80 °C. SERBP1 constructs were purified as previously described for SERBP1 189-400 (Baudin et al, 2021). Briefly, frozen E. coli pellets were thawed and resuspended in 8 M urea, 50 mM Tris pH 8.0, 150 mM NaCl, 20 mM imidazole, lysed by sonication (6 cycles of 10 s on, 30 s off), and the lysate was cleared by centrifugation for 30 min at 45000 g at 4 °C.…”

Section: Protein Expression and Purificationmentioning

confidence: 99%

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Structural Characterization of the RNA-Binding Protein SERBP1 Reveals Intrinsic Disorder and Atypical RNA Binding Modes

Baudin

Moreno-Romero

et al. 2021

Front. Mol. Biosci.

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RNA binding proteins (RBPs) are essential for critical biological processes such as translation regulation and mRNA processing, and misfunctions of these proteins are associated with diseases such as cancer and neurodegeneration. SERBP1 (SERPINE1 mRNA Binding Protein 1) is an RBP that comprises two RG/RGG repeat regions yet lacks other recognizable RNA-binding motifs. It is involved in mRNA maturation, and translational regulation. It was initially identified as a hyaluronic acid binding protein, but recent studies have identified central roles for SERBP1 in brain function and development, especially neurogenesis and synaptogenesis. SERBP1 regulates One-carbon metabolism and epigenetic modification of histones, and increased SERBP1 expression in cancers such as leukemia, ovarian, prostate, liver and glioblastoma is correlated with poor patient outcomes. Despite these important regulatory roles for SERBP1, little is known about its structural and dynamic properties, nor about the molecular mechanisms governing its interaction with mRNA. Here, we define SERBP1 as an intrinsically disordered protein, containing highly conserved elements that were shown to be functionally important. The RNA binding activity of SERBP1 was explored using solution NMR and other biophysical techniques. The outcome of these experiments revealed that SERBP1 preferentially samples compact conformations including a central, stable α-helix and show that SERBP1 recognizes G-rich RNA sequences at the C-terminus involving the RGG box and neighboring residues. Despite the role in RNA recognition, the RGG boxes do not seem to stabilize the central helix and the central helix does not participate in RNA binding. Further, SERBP1 undergoes liquid-liquid phase separation, mediated by salt and RNA, and both RGG boxes are necessary for the efficient formation of condensed phases. Together, these results provide a foundation for understanding the molecular mechanisms of SERBP1 functions in physiological and pathological processes.

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Section: Protein Expression and Purificationmentioning

confidence: 99%

Section: Protein Expression and Purificationmentioning

confidence: 99%

Structural Characterization of the RNA-Binding Protein SERBP1 Reveals Intrinsic Disorder and Atypical RNA Binding Modes

Baudin

Moreno-Romero

et al. 2021

Front. Mol. Biosci.

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Cytosolic RGG RNA-binding proteins are temperature sensitive flowering time regulators in Arabidopsis

Bleckmann,

Spitzlberger,

Denninger

et al. 2023

Biological Chemistry

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mRNA translation is tightly regulated by various classes of RNA-binding proteins (RBPs) during development and in response to changing environmental conditions. In this study, we characterize the arginine-glycine-glycine (RGG) motif containing RBP family of Arabidopsis thaliana representing homologues of the multifunctional translation regulators and ribosomal preservation factors Stm1 from yeast (ScStm1) and human SERBP1 (HsSERBP1). The Arabidopsis genome encodes three RGG proteins named AtRGGA, AtRGGB and AtRGGC. While AtRGGA is ubiquitously expressed, AtRGGB and AtRGGC are enriched in dividing cells. All AtRGGs localize almost exclusively to the cytoplasm and bind with high affinity to ssRNA, while being capable to interact with most nucleic acids, except dsRNA. A protein-interactome study shows that AtRGGs interact with ribosomal proteins and proteins involved in RNA processing and transport. In contrast to ScStm1, AtRGGs are enriched in ribosome-free fractions in polysome profiles, suggesting additional plant-specific functions. Mutant studies show that AtRGG proteins differentially regulate flowering time, with a distinct and complex temperature dependency for each AtRGG protein. In conclusion, we suggest that AtRGGs function in fine-tuning translation efficiency to control flowering time and potentially other developmental processes in response to environmental changes.

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The 1H, 15N and 13C resonance assignments of the C-terminal domain of Serpine mRNA binding protein 1 (SERBP1)

Cited by 2 publications

References 25 publications

Structural Characterization of the RNA-Binding Protein SERBP1 Reveals Intrinsic Disorder and Atypical RNA Binding Modes

Structural Characterization of the RNA-Binding Protein SERBP1 Reveals Intrinsic Disorder and Atypical RNA Binding Modes

Cytosolic RGG RNA-binding proteins are temperature sensitive flowering time regulators in Arabidopsis

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