title: Archaeal RNA ligase from thermoccocus kodakarensis for template dependent ligation.
abstract: Nicking-sealing RNA ligases play a significant biological role in host defense and cellular repair, and have become an important molecular tool in biomedical engineering. Due to the propensity for RNA to form secondary structures, RNA modifying enzymes with elevated optimum temperatures are highly desired. Current characterized double stranded RNA ligases, such as the bacteriophage T4 RNA ligase 2, while possessing good template dependency, are not active at elevated temperatures. The few characterized RNA ligases from thermophiles exhibit high template independency. We synthesize and characterize here, KOD RNA ligase (KOD1Rnl), a thermostable and template dependent RNA ligase from the archaeon, Thermoccocus Kodakarensis. We disclose that a 13 time reduction in template independent ligation can be achieved with the addition of a single stranded DNase, such as RecJ. We also elucidate the effects of the presence of blood proteins on the activity of KOD1Rnl. Template dependent and thermostable RNA ligases, such as KOD RNA ligase, can be utilized in RNA detection, modification and sequencing.
authors.affiliation: a Center for Biomedical Engineering, School of Engineering, Brown University , Providence , RI , USA.
authors.forename: Lei
authors.initials: L
authors.lastname: Zhang
authors.affiliation: a Center for Biomedical Engineering, School of Engineering, Brown University , Providence , RI , USA.
authors.forename: Anubhav
authors.initials: A
authors.lastname: Tripathi
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D029043:Ebolavirus / Q000235:genetics; D008958:Models, Molecular; D011487:Protein Conformation; D011130:RNA Ligase (ATP) / Q000737:chemistry / Q000378:metabolism*; D012367:RNA, Viral / Q000235:genetics; D013379:Substrate Specificity; D013698:Templates, Genetic; D019712:Thermococcus / Q000201:enzymology*
chemical_list: D012367:RNA, Viral; D011130:RNA Ligase (ATP)
keywords: Double-stranded ligase; KOD RNA ligase; RNA sequencing; template dependent ligation; thermostable RNA ligase