6.5.1.3 additional information enzyme does not discriminate between RNA and DNA for phosphodiester bond formation Methanothermobacter thermautotrophicus ? - ? 89 6.5.1.3 additional information intramolecular ligation of 5'-PO4 single-strand RNA to form a covalently closed circular RNA molecule through ligase-adenylylate and RNA-adenylylate intermediates AppRNA. At the optimal temperature of 65°C, AppRNA is predominantly ligated to a circular product. At 35°C, phosphodiester bond formation is suppressed and the majority of the AppRNA is deadenylylated Methanothermobacter thermautotrophicus ? - ? 89 6.5.1.3 additional information T4 RNA ligase possesses nick-joining activity in RNA/DNA hybrids against junctional matches/mismatches complementary to the RNA strand as a ligation template. When a junctional mismatch is present at the 30-terminal of RNA probe, the nick-joining activity of T4Rnl2 is inhibited Tequatrovirus T4 ? - ? 89 6.5.1.3 additional information the RNA ligase has a a strict requirement for RNA substrates with a 2'-phosphate terminus for the ligation of RNAs with 5'-phosphate and 3'-hydroxyl ends. RNA ligase forms a 2'-phosphomonoester-3',5'-phosphodiester junction Branchiostoma floridae ? - ? 89 6.5.1.3 additional information wheat RNA ligase can be dissected into three isolated domain enzymes that are responsible for its core ligase, 5'-kinase, and 2',3'-cyclic phosphate 3'-phosphodiesterase activities, respectively. A side reaction on 5'-tri/diphosphate RNAs is dependent on ATP, a 2'-phosphate-3'-hydroxyl end, and the ligase domain. Two RNA molecules having 5'-hydroxyl and 2',3'-cyclic monophosphate groups are ligated almost stoichiometrically after separate conversion of respective terminal phosphate states into reactive ones Triticum aestivum ? - ? 89 6.5.1.3 additional information wheat RNA ligase contains 5'-hydroxyl kinase, 2',3'-cyclic phosphate 3'-phosphodiesterase, and 5'-phosphate 2'-phosphate-3'-hydroxyl RNA ligase activities Triticum aestivum ? - ? 89 6.5.1.3 additional information RtcA is able of cyclizing a 2'-phosphate RNA end in high yield. The rate of 2'-phosphate cyclization by RtcA is five orders of magnitude slower than 3'-phosphate cyclization, notwithstanding that RtcA binds with similar affinity to RNA3'p and RNA2'p substrates Escherichia coli ? - ? 89 6.5.1.3 additional information the enzyme catalyzes an adenylation reaction that is specific for ATP and either ssDNA or RNA. It does not require synthesis of a template strand for adenylation. To achieve quantitative DNA adenylation, single turnover reaction condition with substrate to enzyme ratio 1:1 should be used Methanothermobacter thermautotrophicus ? - ? 89 6.5.1.3 additional information the enzyme catalyzes formation of phosphodiester bonds between the 5'-phosphate and 3'-hydroxyl termini of single-stranded RNAs. It can also react with RNA with a 3'-phosphate end to generate a 2',3'-cyclic phosphate. The enzyme can additionally remove adenosine from the 3'-terminus of the RNA to produce 3'-deadenylated RNA Methanothermobacter thermautotrophicus ? - ? 89 6.5.1.3 additional information the enzyme is unreactive with a 5'-32P-labeled single-strand 18-mer pDNA substrate Naegleria gruberi ? - ? 89