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Literature summary for 2.7.11.23 extracted from

  • Muehlbacher, W.; Mayer, A.; Sun, M.; Remmert, M.; Cheung, A.C.; Niesser, J.; Soeding, J.; Cramer, P.
    Structure of Ctk3, a subunit of the RNA polymerase II CTD kinase complex, reveals a noncanonical CTD-interacting domain fold (2015), Proteins, 83, 1849-1858.
    View publication on PubMed
Search for more literature for 2.7.11.23

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of C-terminally His6-tagged Ctk3 (residues 1-218) and the Ctk3 N-terminal domain (residues 1-140) in Escherichia coli strain BL21(DE3)RIL or in strain B834 for the selenomethionine-labeled variants Schizosaccharomyces pombe

Crystallization (Commentary)

Crystallization (Comment) Organism
N-terminal domain of the Ctk3 homologue, hanging drop vapor diffusion method, mixing of 14.5 mg/ml protein in 50 mM HEPES pH 8.0, 50 mM NaCl, 1 mM DTT, with reservoir solution containing 26% PEG 6000, 100 mM citric acid, pH 4.0, 0.8 M lithium chloride, and 5 mM Tris(2-carboxyethyl)phosphin, 4°C, X-ray diffraction structure determination and analysis at 2.0 A resolution Schizosaccharomyces pombe

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Schizosaccharomyces pombe

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + [DNA-directed RNA polymerase] Schizosaccharomyces pombe
-
 ADP + phospho-[DNA-directed RNA polymerase]
-
 ?

Organism

Organism UniProt Comment Textmining
Schizosaccharomyces pombe Q9USJ8
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant C-terminally His6-tagged wildtype and selenomethionine-labeled Ctk3 and the Ctk3 N-terminal domain from Escherichia coli strain BL21(DE3)RIL or strain B834 by nickel affinity chromatography, dialysis, tag removal by thrombin cleavage, followed by anion exchange chromatography and gel filtration Schizosaccharomyces pombe

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + [DNA-directed RNA polymerase]
-
 Schizosaccharomyces pombe ADP + phospho-[DNA-directed RNA polymerase]
-
 ?
additional information Ctk3 shows no binding to CTD peptides Schizosaccharomyces pombe ?
-
 ?

Subunits

Subunits Comment Organism
More the enzyme Ctk3 consists of a N-terminal CTD-interacting domain (CID) and a C-terminal three helix bundle domain. The structure reveals eight helices arranged into a right-handed superhelical fold. Ctk3 shows no binding to CTD peptides. Structure and function of the CTDK-I complex, overview. Ctk1 (Lsk1) associates with its cyclin partner Ctk2 (Lsc1) and a third subunit, Ctk3 (Lsg1), to form the CTD kinase I (CTDK-I) complex. This trimeric structure is unique amongst CDK complexes. The Ctk3 N-terminal domain has a noncanonical surface Schizosaccharomyces pombe

Synonyms

Synonyms Comment Organism
CTD kinase
-
 Schizosaccharomyces pombe
CTD kinase subunit 3
-
 Schizosaccharomyces pombe
CTD kinase subunit gamma
-
 Schizosaccharomyces pombe
CTDK-I
-
 Schizosaccharomyces pombe
CTDK-I gamma subunit
-
 Schizosaccharomyces pombe
Ctk3
-
 Schizosaccharomyces pombe
RNA polymerase II CTD kinase
-
 Schizosaccharomyces pombe

Cofactor

Cofactor Comment Organism Structure
ATP
-
 Schizosaccharomyces pombe

General Information

General Information Comment Organism
additional information structure and function of the CTDK-I complex, overview. Prediction of a possible CTD-binding domain (CID) in the N-terminal region of Ctk3 and of a three-helix bundle in the C-terminal region of Ctk3 Schizosaccharomyces pombe
physiological function CTDK-I is a yeast kinase complex that phosphorylates the C-terminal repeat domain (CTD) of RNA polymerase II (Pol II) to promote transcription elongation. CTDK-I contains the cyclin-dependent kinase Ctk1 (homologous to human CDK9/CDK12), the cyclin Ctk2 (human cyclin K), and the yeast-specific subunit Ctk3, which is required for CTDK-I stability and activity Schizosaccharomyces pombe