ArticleNucleic acids research2014
Efficient DNA ligation in DNA-RNA hybrid helices by Chlorella virus DNA ligase.
Article in Nucleic acids research, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 47 papers.
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Who cites it
47 citing papers in PubMed, 111 citations in OpenAlex.
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- Advances in nucleic acid probe-based detection of gene point mutations: a review.Frontiers in chemistry · 2025Review
- Template-dependent DNA ligation for the synthesis of modified oligonucleotides.Nature communications · 2024Article
- Spatial multi-omics at subcellular resolution via high-throughput in situ pairwise sequencing.Nature biomedical engineering · 2024Article
- Characterisation and engineering of a thermophilic RNA ligase from Palaeococcus pacificus.Nucleic acids research · 2024Article
- Single-Cell Imaging of mRNA by Target RNA-Initiated RCA.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Viral replication organelles: the highly complex and programmed replication machinery.Frontiers in microbiology · 2024Review
- Detection of viral RNAs at ambient temperature via reporter proteins produced through the target-splinted ligation of DNA probes.Nature biomedical engineering · 2023Article
- A one-pot isothermal Cas12-based assay for the sensitive detection of microRNAs.Nature biomedical engineering · 2023Article
- Mesoporous Polydopamine-Encapsulated Fluorescent Nanodiamonds: A Versatile Platform for Biomedical Applications.ACS applied materials & interfaces · 2023Article
- The importance of mRNA (New York, N.Y.) · 2023Article
- RT-IVT method allows multiplex real-time quantification of in vitro transcriptional mRNA production.Communications biology · 2023Article
- Quantitative analysis of miRNAs using SplintR ligase-mediated ligation of complementary-pairing probes enhanced by RNase H (SPLICER)-qPCR.Molecular therapy. Nucleic acids · 2023Article
- Hierarchical DNA branch assembly-encoded fluorescent nanoladders for single-cell transcripts imaging.Nucleic acids research · 2023Article
- Giant Viruses as a Source of Novel Enzymes for Biotechnological Application.Pathogens (Basel, Switzerland) · 2022Review
- Direct RNA targeted in situ sequencing for transcriptomic profiling in tissue.Scientific reports · 2022Article
Corrections and comments
- Erratum issued
Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Single-stranded DNA molecules (ssDNA) annealed to an RNA splint are notoriously poor substrates for DNA ligases. Herein we report the unexpectedly efficient ligation of RNA-splinted DNA by Chlorella virus DNA ligase (PBCV-1 DNA ligase). PBCV-1 DNA ligase ligated ssDNA splinted by RNA with kcat ≈ 8 x 10(-3) s(-1) and K(M) < 1 nM at 25 °C under conditions where T4 DNA ligase produced only 5'-adenylylated DNA with a 20-fold lower kcat and a K(M) ≈ 300 nM. The rate of ligation increased with addition of Mn(2+), but was strongly inhibited by concentrations of NaCl >100 mM. Abortive adenylylation was suppressed at low ATP concentrations (<100 µM) and pH >8, leading to increased product yields. The ligation reaction was rapid for a broad range of substrate sequences, but was relatively slower for substrates with a 5'-phosphorylated dC or dG residue on the 3' side of the ligation junction. Nevertheless, PBCV-1 DNA ligase ligated all sequences tested with 10-fold less enzyme and 15-fold shorter incubation times than required when using T4 DNA ligase. Furthermore, this ligase was used in a ligation-based detection assay system to show increased sensitivity over T4 DNA ligase in the specific detection of a target mRNA.
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