DNA molecules are a promising data storage medium for the future; however, effective synthesis of DNA using an enzyme that catalyzes the polymerization of natural nucleoside triphosphates in a user-defined manner, without the need for multiple injections of polymerase, remains a challenge. In the present study, we demonstrated that the bacteriophage abortive infection system reverse transcriptase AbiK from facilitates such an approach. We employed surface plasmon resonance to monitor the polymerization of the DNA strand with a user-defined sequence of multiple segments through a sequential buffer exchange process.
View Article and Find Full Text PDFReverse transcriptases (RTs) are enzymes with DNA polymerase and RNase H activities. They convert ssRNA into dsDNA and are key enzymes for the replication of retroviruses and retroelements. Caulimoviridae is a major family of plant-infecting viruses.
View Article and Find Full Text PDFBacterial reverse transcriptases (RTs) are a large and diverse enzyme family. AbiA, AbiK and Abi-P2 are abortive infection system (Abi) RTs that mediate defense against bacteriophages. What sets Abi RTs apart from other RT enzymes is their ability to synthesize long DNA products of random sequences in a template- and primer-independent manner.
View Article and Find Full Text PDFCoronaviruses modify their single-stranded RNA genome with a methylated cap during replication to mimic the eukaryotic mRNAs. The capping process is initiated by several nonstructural proteins (nsp) encoded in the viral genome. The methylation is performed by two methyltransferases, nsp14 and nsp16, while nsp10 acts as a co-factor to both.
View Article and Find Full Text PDFCoronaviruses contain one of the largest genomes among the RNA viruses, coding for 14-16 non-structural proteins (nsp) that are involved in proteolytic processing, genome replication and transcription, and four structural proteins that build the core of the mature virion. Due to conservation across coronaviruses, nsps form a group of promising drug targets as their inhibition directly affects viral replication and, therefore, progression of infection. A minimal but fully functional replication and transcription complex was shown to be formed by one RNA-dependent RNA polymerase (nsp12), one nsp7, two nsp8 accessory subunits, and two helicase (nsp13) enzymes.
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