4XPZ

Structure of fission yeast RNA polymerase II CTD phosphatase Fcp1-R271A bound to aluminum fluoride

Summary for 4XPZ

• Entry DOI: 10.2210/pdb4xpz/pdb
• Related: 3EF0 3EF1 4XQ0
• Descriptor: RNA polymerase II subunit A C-terminal domain phosphatase, ALUMINUM FLUORIDE, TETRAETHYLENE GLYCOL, ... (5 entities in total)
• Functional Keywords: phosphatase, polii-ctd, transition state, alf3, hydrolase
• Biological source: Schizosaccharomyces pombe (strain 972 / ATCC 24843) (Fission yeast); More
• Total number of polymer chains: 1
• Total formula weight: 42794.05

Authors

Ghosh, A.,Lima, C.D. (deposition date: 2015-01-18, release date: 2015-03-04, Last modification date: 2023-09-27)

Primary citation

Schwer, B.,Ghosh, A.,Sanchez, A.M.,Lima, C.D.,Shuman, S.
Genetic and structural analysis of the essential fission yeast RNA polymerase II CTD phosphatase Fcp1.
Rna, 21:1135-1146, 2015

PubMed Abstract: Protein phosphatases regulate mRNA synthesis and processing by remodeling the carboxy-terminal domain (CTD) of RNA polymerase II (Pol2) to dynamically inscribe a Pol2 CTD code. Fission yeast Fcp1 (SpFcp1) is an essential 723-amino acid CTD phosphatase that preferentially hydrolyzes Ser2-PO4 of the YS(2)PTSPS repeat. The SpFcp1 catalytic domain (aa 140-580) is composed of a DxDxT acyl-phosphatase module (FCPH) and a BRCT module. Here we conducted a genetic analysis of SpFcp1, which shows that (i) phosphatase catalytic activity is required for vegetative growth of fission yeast; (ii) the flanking amino-terminal domain (aa 1-139) and its putative metal-binding motif C(99)H(101)Cys(109)C(112) are essential; (iii) the carboxy-terminal domain (aa 581-723) is dispensable; (iv) a structurally disordered internal segment of the FCPH domain (aa 330-393) is dispensable; (v) lethal SpFcp1 mutations R271A and R299A are rescued by shortening the Pol2 CTD repeat array; and (vi) CTD Ser2-PO4 is not the only essential target of SpFcp1 in vivo. Recent studies highlight a second CTD code involving threonine phosphorylation of a repeat motif in transcription elongation factor Spt5. We find that Fcp1 can dephosphorylate Thr1-PO4 of the fission yeast Spt5 CTD nonamer repeat T(1)PAWNSGSK. We identify Arg271 as a governor of Pol2 versus Spt5 CTD substrate preference. Our findings implicate Fcp1 as a versatile sculptor of both the Pol2 and Spt5 CTD codes. Finally, we report a new 1.45 Å crystal structure of SpFcp1 with Mg(2+) and AlF3 that mimics an associative phosphorane transition state of the enzyme-aspartyl-phosphate hydrolysis reaction.

PubMed: 25883047
DOI: 10.1261/rna.050286.115

Experimental method

X-RAY DIFFRACTION (1.45 Å)