Biophysical and biochemical analysis of hnRNP K: arginine methylation, reversible aggregation and combinatorial binding to nucleic acids

Moritz, Bodo; Lilie, Hauke; Vries, Isabel S. Naarmann-de; Urlaub, Henning; Wahle, Elmar; Ostareck-Lederer, Antje; Ostareck, Dirk H.

doi:10.1515/hsz-2014-0146

Cited by 16 publications

(18 citation statements)

References 76 publications

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“…For this, we expressed the PRMT1 methyltransferase and the target protein from the same plasmid in E. coli ( Fig. 4C; Moritz et al 2014) and purified AUF1 p45 aDMA to homogeneity using the analogous purification protocol as earlier with the nonmethylated AUF1 p45 ( Fig. 4D; Friedrich et al 2014).…”

Section: Auf1 P45 Is Dimethylated At Five Arginine Residues In the C mentioning

confidence: 99%

Arginine methylation enhances the RNA chaperone activity of the West Nile virus host factor AUF1 p45

Friedrich

Schmidt

Schierhorn

et al. 2016

RNA

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A prerequisite for the intracellular replication process of the Flavivirus West Nile virus (WNV) is the cyclization of the viral RNA genome, which enables the viral replicase to initiate RNA synthesis. Our earlier studies indicated that the p45 isoform of the cellular AU-rich element binding protein 1 (AUF1) has an RNA chaperone activity, which supports RNA cyclization and viral RNA synthesis by destabilizing a stem structure at the WNV RNA's 3 ′ -end. Here we show that in mammalian cells, AUF1 p45 is consistently modified by arginine methylation of its C terminus. By a combination of different experimental approaches, we can demonstrate that the methyltransferase PRMT1 is necessary and sufficient for AUF1 p45 methylation and that PRMT1 is required for efficient WNV replication. Interestingly, in comparison to the nonmethylated AUF1 p45, the methylated AUF1 p45 aDMA exhibits a significantly increased affinity to the WNV RNA termini. Further data also revealed that the RNA chaperone activity of AUF1 p45 aDMA is improved and the methylated protein stimulates viral RNA synthesis considerably more efficiently than the nonmethylated AUF1 p45. In addition to its destabilizing RNA chaperone activity, we identified an RNA annealing activity of AUF1 p45, which is not affected by methylation. Arginine methylation of AUF1 p45 thus represents a specific determinant of its RNA chaperone activity while functioning as a WNV host factor. Our data suggest that the methylation modifies the conformation of AUF1 p45 and in this way affects its RNA binding and restructuring activities.

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Section: Auf1 P45 Is Dimethylated At Five Arginine Residues In the C mentioning

confidence: 99%

Arginine methylation enhances the RNA chaperone activity of the West Nile virus host factor AUF1 p45

Friedrich

Schmidt

Schierhorn

et al. 2016

RNA

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“…It is active at the chromatin level, and present in greater density near transcribed genes with respect to silent ones. hnRNP K directly binds to the promoter region of the human c-myc gene (23) and was found to promote neoplastic transformation in an eIf4E-dependent manner (24). The expression level of hnRNP K was higher in melanoma, breast and prostate cancers than that in normal control groups (14,25,26).…”

Section: Discussionmentioning

confidence: 99%

Heterogeneous nuclear ribonucleoprotein K is overexpressed and associated with poor prognosis in gastric cancer

Yang

Zeng

et al. 2016

Oncology Reports

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Heterogeneous nuclear ribonucleoprotein K (hnRNP K) is one of the major pre-mRNA-binding proteins, that is involved in translational modifications. In our previous studies, we found that hnRNP K is associated with human gastric cancer. The protein levels of hnRNP K were detected in cell lines and tissue microarrays. The correlation between hnRNP K expression and patient survival rate was evaluated by Kaplan-Meier survival analysis. In addition, we also detected hnRNP K expression in preoperative and postoperative serum samples from patients with gastric cancer, and serum samples from healthy volunteers. We found that hnRNP K was overexpressed in the gastric cancer cell lines. The levels of hnRNP K were significantly elevated in the gastric cancer tissues compared with that noted in the tumor-adjacent gastric mucosal and normal gastric mucosal sampes, and hnRNP K expression was found to correlate with tumor stage and lymph node metastasis. However, the level of serum hnRNP K did not differ significantly between gastric cancer patients and healthy volunteers. We also found that patients whose tumors showed elevated expression of hnRNP K had poor survival. The present study suggests that hnRNP K is a promising tissue biomarker for diagnosing gastric cancer and is a prognostic indicator for patients with gastric cancer.

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“…2A), combined with microarray analysis revealed 155 mRNAs to be hnRNP-K-enriched in non-induced cells that were classified by PANTHER (Mi et al, 2013;Thomas et al, 2003) and gene ontology domain annotation (Tables S1-S4). Functional relevant hnRNP-KmRNA interactions were determined based on criteria including biochemical, biophysical and structural analyses of hnRNP K interaction sequences (Backe et al, 2005;Messias et al, 2006;Moritz et al, 2014;Ostareck et al, 1997). Initially applying the hnRNP-K-binding motif (two U/CCCC motifs within 19 nts) as a criterion, 113 mRNAs representing 64% of the isolated population were identified ( Fig.…”

Section: Discussionmentioning

confidence: 99%

“…To classify all identified protein coding mRNAs, PANTHER protein class annotation (Mi et al, 2013;Thomas et al, 2003) (Table S1) and gene ontology domain assignment to molecular function, biological process and cellular component (Tables S2-S4) were applied. As a further criterion for the specific enrichment of identified mRNAs, we used well-characterized RNA sequence characteristics (Backe et al, 2005;Messias et al, 2006;Moritz et al, 2014;Ostareck et al, 1997): within a sequence range of 19 nts at least two U/CCCC motifs are sufficient for hnRNP K binding. Of the 155 identified mRNAs 64% (113) fulfil that criterion.…”

Section: Differential Hnrnp-k−rna Binding In Erythroid Maturationmentioning

confidence: 99%

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Translational control mediated by hnRNP K links NMHC IIA to erythroid enucleation

Vries

Brendle

Bähr-Ivacevic

et al. 2016

Journal of Cell Science

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Post-transcriptional regulation is crucial for structural and functional alterations in erythropoiesis. Enucleation of erythroid progenitors precedes reticulocyte release into circulation. In enucleated cells, reticulocyte 15-lipoxygenase (r15-LOX, also known as ALOX15) initiates mitochondria degradation. Regulation of r15-LOX mRNA translation by hnRNP K determines timely r15-LOX synthesis in terminal maturation. K562 cells induced for erythroid maturation recapitulate enucleation and mitochondria degradation. HnRNP K depletion from maturing K562 cells results in enhanced enucleation, which even occurs independently of maturation. We performed RIP-Chip analysis to identify hnRNP K-interacting RNAs comprehensively. Non-muscle myosin heavy chain (NMHC) IIA (also known as MYH9) mRNA co-purified with hnRNP K from noninduced K562 cells, but not from mature cells. NMHC IIA protein increase in erythroid maturation at constant NMHC IIA mRNA levels indicates post-transcriptional regulation. We demonstrate that binding of hnRNP K KH domain 3 to a specific sequence element in the NMHC IIA mRNA 3′UTR mediates translation regulation in vitro. Importantly, elevated NMHC IIA expression results in erythroidmaturation-independent enucleation as shown for hnRNP K depletion. Our data provide evidence that hnRNP-K-mediated regulation of NMHC IIA mRNA translation contributes to the control of enucleation in erythropoiesis.

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Biophysical and biochemical analysis of hnRNP K: arginine methylation, reversible aggregation and combinatorial binding to nucleic acids

Cited by 16 publications

References 76 publications

Arginine methylation enhances the RNA chaperone activity of the West Nile virus host factor AUF1 p45

Arginine methylation enhances the RNA chaperone activity of the West Nile virus host factor AUF1 p45

Heterogeneous nuclear ribonucleoprotein K is overexpressed and associated with poor prognosis in gastric cancer

Translational control mediated by hnRNP K links NMHC IIA to erythroid enucleation

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