Protein kinase CK2 interacts with the splicing factor hPrp3p

Lehnert, S. A.; Götz, Claudia; Kartarius, Sabine; Schäfer, Barbara Nicole; Montenarh, Mathias

doi:10.1038/sj.onc.1210882

Cited by 20 publications

(15 citation statements)

References 35 publications

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“…It was previously demonstrated that CK2 phosphorylates SNAPC4 and this phosphorylation leads to loss in SNAPC4 DNA binding and loss of U6 transcriptional activity (Gu et al, 2007 ). A number of general splicing factors were also identified, such as SLU7, DHX16, and MFAP1, which again agrees with a general role for CK2 in mRNA splicing (Lehnert et al, 2008 ). Another cluster of tightly-associated interactors (red) included proteins involved in cell division, for instance members of the condensin I complex including (NCAPD2 and NCAPH).…”

Section: Resultssupporting

confidence: 73%

Identification of Candidate Casein Kinase 2 Substrates in Mitosis by Quantitative Phosphoproteomics

Rusin

Adamo

Kettenbach

2017

Front. Cell Dev. Biol.

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Protein phosphorylation is a crucial regulatory mechanism that controls many aspects of cellular signaling. Casein kinase 2 (CK2), a constitutively expressed and active kinase, plays key roles in an array of cellular events including transcription and translation, ribosome biogenesis, cell cycle progression, and apoptosis. CK2 is implicated in cancerous transformation and is a therapeutic target in anti-cancer therapy. The specific and selective CK2 ATP competitive inhibitor, CX-4945 (silmitaseratib), is currently in phase 2 clinical trials. While many substrates and interactors of CK2 have been identified, less is known about CK2 substrates in mitosis. In the present work, we utilize CX-4945 and quantitative phosphoproteomics to inhibit CK2 activity in mitotically arrested HeLa cells and determine candidate CK2 substrates. We identify 330 phosphorylation sites on 202 proteins as significantly decreased in abundance upon inhibition of CK2 activity. Motif analysis of decreased sites reveals a linear kinase motif with aspartic and glutamic amino acids downstream of the phosphorylated residues, which is consistent with known substrate preferences for CK2. To validate specific candidate CK2 substrates, we perform in vitro kinase assays using purified components. Furthermore, we identified CK2 interacting proteins by affinity purification-mass spectrometry (AP-MS). To investigate the biological processes regulated by CK2 in mitosis, we perform network analysis and identify an enrichment of proteins involved in chromosome condensation, chromatin organization, and RNA processing. We demonstrate that overexpression of CK2 in HeLa cells affects proper chromosome condensation. Previously, we found that phosphoprotein phosphatase 6 (PP6), but not phosphoprotein phosphatase 2A (PP2A), opposes CK2 phosphorylation of the condensin I complex, which is essential for chromosome condensation. Here, we extend this observation and demonstrate that PP6 opposition of CK2 is a more general cellular regulatory mechanism.

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Section: Resultssupporting

confidence: 73%

Identification of Candidate Casein Kinase 2 Substrates in Mitosis by Quantitative Phosphoproteomics

Rusin

Adamo

Kettenbach

2017

Front. Cell Dev. Biol.

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“…It has previously been shown that SKIP/SWN1 plays independent roles in splicing and transcription elongation [45]. KIN-3 encodes the ortholog of the human catalytic subunit of CKIIα, which is a protein kinase that phosphorylates several substrates regulating transcription and physically interacts with the splicing factor PRPF3 [46]. Finally, PRP-19 is the ortholog of human PRPF19, and the conserved Prp19 complex (Prp19C) is involved in pre-mRNA processing and transcriptional efficiency [13].…”

Section: Resultsmentioning

confidence: 99%

RSR-2, the Caenorhabditis elegans Ortholog of Human Spliceosomal Component SRm300/SRRM2, Regulates Development by Influencing the Transcriptional Machinery

et al. 2013

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Protein components of the spliceosome are highly conserved in eukaryotes and can influence several steps of the gene expression process. RSR-2, the Caenorhabditis elegans ortholog of the human spliceosomal protein SRm300/SRRM2, is essential for viability, in contrast to the yeast ortholog Cwc21p. We took advantage of mutants and RNA interference (RNAi) to study rsr-2 functions in C. elegans, and through genetic epistasis analysis found that rsr-2 is within the germline sex determination pathway. Intriguingly, transcriptome analyses of rsr-2(RNAi) animals did not reveal appreciable splicing defects but instead a slight global decrease in transcript levels. We further investigated this effect in transcription and observed that RSR-2 colocalizes with DNA in germline nuclei and coprecipitates with chromatin, displaying a ChIP-Seq profile similar to that obtained for the RNA Polymerase II (RNAPII). Consistent with a novel transcription function we demonstrate that the recruitment of RSR-2 to chromatin is splicing-independent and that RSR-2 interacts with RNAPII and affects RNAPII phosphorylation states. Proteomic analyses identified proteins associated with RSR-2 that are involved in different gene expression steps, including RNA metabolism and transcription with PRP-8 and PRP-19 being the strongest interacting partners. PRP-8 is a core component of the spliceosome and PRP-19 is the core component of the PRP19 complex, which interacts with RNAPII and is necessary for full transcriptional activity. Taken together, our study proposes that RSR-2 is a multifunctional protein whose role in transcription influences C. elegans development.

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“…Recombinant CK2 holoenzyme cloned in a bicistronic vector [26] and the three different subunits (a, a 0 , and b in pT7-7) of protein kinase CK2 were expressed in Escherichia coli BL21 (DE3) and the proteins were purified according to published protocols [27,28].…”

Section: Purification Of Ck2 Subunits and Holoenzymementioning

confidence: 99%

Protein kinase CK2 and new binding partners during spermatogenesis

Mannowetz

Kartarius

Wennemuth

et al. 2010

Cell. Mol. Life Sci.

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Protein kinase CK2 is an ubiquitously expressed enzyme that is absolutely necessary for the survival of cells. Besides the holoenzyme consisting of the regulatory β-subunit and the catalytic α- or α'-subunit, the subunits exist in separate forms. The subunits bind to a number of other cellular proteins. We show the expression of individual subunits as well as interaction with the transitional nuclear protein TNP1 and with the motor neuron protein KIF5C during spermatogenesis. TNP1 is a newly identified binding partner of the α-subunit of CK2. CK2α and KIF5C were found in late spermatogenesis, whereas CK2β and TNP1 were found in early spermatogenesis. CK2α, CK2α', TNP1, and KIF5C were detected in the acrosome of spermatozoa, while CK2β was detectable in the mid-piece. Combinations of CK2 subunits might determine interactions with other proteins during spermatogenesis. KIF5C as a kinesin motor neuron protein is probably involved in the redistribution of proteins during spermatogenesis.

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Protein kinase CK2 interacts with the splicing factor hPrp3p

Cited by 20 publications

References 35 publications

Identification of Candidate Casein Kinase 2 Substrates in Mitosis by Quantitative Phosphoproteomics

Identification of Candidate Casein Kinase 2 Substrates in Mitosis by Quantitative Phosphoproteomics

RSR-2, the Caenorhabditis elegans Ortholog of Human Spliceosomal Component SRm300/SRRM2, Regulates Development by Influencing the Transcriptional Machinery

Protein kinase CK2 and new binding partners during spermatogenesis

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