EC Number   |
|---|
   2.7.7.19 | alone and in complex with 3-dATP |
| 2.7.7.19 | ATP-gamma-S bound and unbound structures. Subunit VP55 residues of the active site make specific interactions with ATP-gamma-S. Concave surface of subunit VP55 docks the globular subunit VP39. Model of RNA primer binding shows that subunit VP39 functions as a processivity factor by partially enclosing the RNA primer at the heterodimer interface |
| 2.7.7.19 | core complex of isoform GLD-2 and RRM RNA binding domain protein RNP-8, to 2.5 A resolution. RNP-8 embraces the poly(A)-polymerase, docking onto several conserved hydrophobic hotspots present on the GLD-2 surface. RNP-8 stabilizes GLD-2 and indirectly stimulates polyadenylation. RNP-8 differs in amino-acid sequence and structure from GLD-2 binding partner GLD-3 but binds the same surfaces of GLD-2 by forming alternative interactions |
| 2.7.7.19 | Crystals of PAP complexed to 3'-dATP and Mn2+ are grown from a solution made of 22% (w/v) polyethylene glycol 8000, 100 mM Mes (pH 6), 120 mM ammonium sulfate, 5 mM CaCl2, 2 mM MnCl2, and 2 mM beta-mercaptoethanol. Cocrystallization of PAP with 3'-dATP and MgCl2 is performed under conditions similar to those reported, with the exception that 5 mM MgCl2 is used instead of 2 mM MnCl2. Crystals are of the space group p212121, with one monomer per asymmetric unit and a solvent content of ~55%. |
| 2.7.7.19 | germ-line development defective GLD-2GLD-3 complex up-regulates the expression of genes required for meiotic progression. The structure of a minimal polyadenylation complex that includes the conserved nucleotidyl-transferase core of GLD-2 and the N-terminal domain of GLD-3, to 2.3 A resolution, shows that the N-terminal domain of GLD-3 does not fold into the predicted multi-K homology domain but wraps around the catalytic domain of GLD-2. GLD-3 activates GLD-2 both indirectly by stabilizing the enzyme and directly by contributing positively charged residues near the RNA-binding cleft. Due to distinct structural features, GLD-2 displays unusual specificity in vitro for single-stranded RNAs with at least one adenosine at the 3'-end |
| 2.7.7.19 | hanging drop vapor diffusion method, using 2% (w/v) tacsimate pH 5.0, 0.15 M sodium citrate tribasic dehydrate pH 5.6 and 10% (w/v) PEG 3350 at 10°C |
| 2.7.7.19 | hanging drop vapour diffusion method, using 20% (w/v) PEG 8000, 100 mM magnesium acetate, 100 mM imidazole (pH 6.2), 3% ethylene glycol |
| 2.7.7.19 | in complex with an ATP analog |
| 2.7.7.19 | isoform PAPgamma bound to cordycepin triphosphate (3?dATP) and Ca2+, to 2.8 A resolution. One 3'-dATP and one Ca2+ are present in the active site of each PAP molecule. Strictly conserved catalytic residues Asp112 and Asp114 interact with Ca2+, which also ligates three non-bridging oxygens of the alpha, beta and gamma phosphates of 3'-dATP. PAPgamma closely resembles its PAPalpha ortholog |
| 2.7.7.19 | mature enzyme, to 1.82 A resolution. Enzyme crystallizes as a dimer and consists of a N-terminal domain (NTD: 52-194 aa) and of palm (195-341 aa) and fingers (342-527 aa). The palm domain harbors the catalytic triad, residue D237, located at the base of sheet beta6, D239 on strand beta6 and D319, which belongs to beta10. Structures of mtPAP in complex with cosubstrates UTP, CTP and GTP indicate that initial nucleotide selection occurs in the absence of a template |
