Abstract
With their ability to catalyse the formation of phosphodiester linkages, DNA ligases and RNA ligases are essential tools for many protocols in molecular biology and biotechnology. Currently, the nucleic acid ligases from bacteriophage T4 are used extensively in these protocols. In this review, we argue that the nucleic acid ligases from Archaea represent a largely untapped pool of enzymes with diverse and potentially favourable properties for new and emerging biotechnological applications. We summarise the current state of knowledge on archaeal DNA and RNA ligases, which makes apparent the relative scarcity of information on in vitro activities that are of most relevance to biotechnologists (such as the ability to join blunt- or cohesive-ended, double-stranded DNA fragments). We highlight the existing biotechnological applications of archaeal DNA ligases and RNA ligases. Finally, we draw attention to recent experiments in which protein engineering was used to modify the activities of the DNA ligase from Pyrococcus furiosus and the RNA ligase from Methanothermobacter thermautotrophicus, thus demonstrating the potential for further work in this area.
Highlights
DNA and RNA ligases are ubiquitous enzymes that catalyse the formation of phosphodiester bonds between opposing 5-phosphate and 3-hydroxyl termini in nucleic acids [1,2,3]
We summarise the current state of knowledge about these enzymes, including their existing applications in biotechnology, and we argue that they offer a largely untapped pool of activities for use in “ generation” molecular biology protocols
While it is yet to be tested with other archaeal DNA ligases, this design approach, releasing the interactions of the C-terminal helix with the adenylation domain (AdD) and oligonucleotide-binding domain (OBD) domains, appears as though it could be a generally applicable one for enhancing activity
Summary
DNA and RNA ligases are ubiquitous enzymes that catalyse the formation of phosphodiester bonds between opposing 5-phosphate and 3-hydroxyl termini in nucleic acids [1,2,3]. Their activities are essential for central biological processes, including DNA replication and recombination, rearrangement of immunoglobulin genes, and RNA editing and repair. Their activities in vitro have been exploited in numerous molecular biology protocols, making them critical tools for modern biotechnology. We summarise the current state of knowledge about these enzymes, including their existing applications in biotechnology, and we argue that they offer a largely untapped pool of activities for use in “ generation” molecular biology protocols
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