An Encephalitozoon cuniculi ortholog of the RNA polymerase II carboxyl-terminal domain (CTD) serine phosphatase Fcp1 Journal Article
| Authors: | Hausmann, S.; Schwer, B.; Shuman, S. |
| Article Title: | An Encephalitozoon cuniculi ortholog of the RNA polymerase II carboxyl-terminal domain (CTD) serine phosphatase Fcp1 |
| Abstract: | Fcp1 is an essential protein serine phosphatase that dephosphorylates Ser2 or Ser5 of the RNA pglymerase II carboxyl-terminal domain (CTD) heptad repeat Y1S2P3T4S5P 6S7. The CTD of the microsporidian parasite Encephalitozoon cuniculi consists of 15 heptad repeats, which approximates the minimal CTD length requirement for cell viability in yeast. Here we show that E. cuniculi encodes a minimized 411-aa Fcp1-like protein (EcFcp1), which consists of a DxDx(T/V) phosphatase domain and a BRCA1 carboxyl terminus (BRCT) domain but lacks the large N- and C-terminal domains found in fungal and metazoan Fcp1 enzymes. Nonetheless, EcFcp1 can function in lieu of Saccharomyces cerevisiae Fcp1 to sustain yeast cell growth. Recombinant EcFcp1 is a monomeric enzyme with intrinsic phosphatase activity against nonspecific (p-nitrophenyl phosphate) and specific (CTD-PO4) substrates. EcFcp1 dephosphorylates CTD positions Ser2 and Ser5 with similar efficacy in vitro. We exploit synthetic CTD Ser2-PO4 and Ser5-PO4 peptides to define minimized substrates for EcFcp1 and to illuminate the importance of CTD primary structure in Ser2 and Ser5 phosphatase activity. |
| Keywords: | unclassified drug; human cell; nonhuman; polymerase chain reaction; protein domain; animals; cell viability; cell division; cell growth; phosphatase; serine; enzyme activity; enzyme substrate; phosphorylation; brca1 protein; rna; amino acid sequence; molecular sequence data; sequence homology, amino acid; saccharomyces cerevisiae; nucleotide sequence; peptide fragments; binding sites; yeast; sequence homology; enzyme subunit; phosphates; rna polymerase ii; enzyme assay; enzymes; yeast cell; polyacrylamide gel electrophoresis; phosphoprotein phosphatase; metazoa; cells; dephosphorylation; saccharomyces; repetitive sequences, amino acid; encephalitozoon cuniculi; carboxyl-terminal domain (ctd); structure (composition); microsporidia; human; priority journal; article; fcp1 like protein; serine phosphatase |
| Journal Title: | Biochemistry |
| Volume: | 43 |
| Issue: | 22 |
| ISSN: | 0006-2960 |
| Publisher: | American Chemical Society |
| Date Published: | 2004-06-08 |
| Start Page: | 7111 |
| End Page: | 7120 |
| Language: | English |
| DOI: | 10.1021/bi0499617 |
| PROVIDER: | scopus |
| PUBMED: | 15170348 |
| DOI/URL: | |
| Notes: | Biochemistry -- Cited By (since 1996):12 -- Export Date: 16 June 2014 -- CODEN: BICHA -- Molecular Sequence Numbers: GENBANK: CAD26175; -- Source: Scopus |
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