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Literature summary for 2.7.7.48 extracted from

  • Ren, Z.; C Franklin, M.; Ghose, R.
    Structure of the RNA-directed RNA polymerase from the cystovirus phi12 (2013), Proteins, 81, 1479-1484.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
recombinant enzyme P2 expression in Escherichia coli strain BL21 (DE3) Pseudomonas phage phi12

Crystallization (Commentary)

Crystallization (Comment) Organism
enzyme in two crystal forms A and B, method screening, 300 nl of 6-20 mg/ml protein in 50 mM Tris, 100 mM NaCl, and 2 mM DTT, pH 8.0, are mixed with reservoir solution containing 4.0 M sodium formate, 0.2 M magnesium formate, and 20% w/v PEG 3350, in hanging drops at 20°C, or with 4.3 M NaCl, 0.1 M HEPES, and 30 mM tri-glycine in sitting drops at room temperature, X-ray diffraction structure determination and analysis at resolutions of 1.7 A and 2.1 A, respectively. Form A contains Mg2+ bound at a site that deviates from the canonical non-catalytic position seen in form B Pseudomonas phage phi12

Protein Variants

Protein Variants Comment Organism
additional information temperature sensitivity of a ts-mutant Pseudomonas phage phi12

Inhibitors

Inhibitors Comment Organism Structure
Ca2+ inhibits catalyis Pseudomonas phage phi12

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required for catalyis Pseudomonas phage phi12
additional information Mn2+ is not required for catalyis Pseudomonas phage phi12

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
nucleoside triphosphate + RNAn Pseudomonas phage phi12
-
diphosphate + RNAn+1
-
?

Organism

Organism UniProt Comment Textmining
Pseudomonas phage phi12 Q94M06
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme P2 from Escherichia coli strain BL21 (DE3) by anion exchange chromatography, gel filtration, dialysis, and ultrafiltration Pseudomonas phage phi12

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information template ssRNA accesses the active site through a tunnel. The phi12 P2 conformation does not favor RNA binding Pseudomonas phage phi12 ?
-
?
nucleoside triphosphate + RNAn
-
Pseudomonas phage phi12 diphosphate + RNAn+1
-
?

Synonyms

Synonyms Comment Organism
RDRP
-
Pseudomonas phage phi12
self-priming RdRp
-
Pseudomonas phage phi12

General Information

General Information Comment Organism
additional information structure of P2, the self-priming RdRp from cystovirus phi12, crystal structure analysis, overview. The tunnel through which template ssRNA accesses the active site is partially occluded by a flexible loop; this feature, along with sub-optimal positioning of other structural elements that prevent the formation of a stable initiation complex, indicate an inactive conformation in crystallo Pseudomonas phage phi12
physiological function P2, an RNA-directed polymerase (RdRp), is the critical component of a four-protein polymerase complex (PX) that includes in addition to P2, the major capsid protein, P1, a packaging NTPase, P4, and an accessory protein, P71. P2 performs the dual tasks of transcription and replication de novo without the use of a primer (i. e. P2 is a self-priming RdRp), and plays a critical role in the cystoviral life cycle Pseudomonas phage phi12