The replicative helicases of bacteria, archaea, and eukarya can unwind RNA-DNA hybrid substrates.
Replicative helicases are hexameric enzymes that unwind DNA during chromosomal replication. They use energy from nucleoside triphosphate hydrolysis to translocate along one strand of the duplex DNA and displace the complementary strand. Here, the ability of a replicative helicase from each of the three domains, bacteria, archaea, and eukarya, to unwind RNA-containing substrate was determined. It is shown that all three helicases can unwind DNA-RNA hybrids while translocating along the single-stranded DNA. No unwinding could be observed when the helicases were provided with a single-stranded RNA overhang. Using DNA, RNA, and DNA-RNA chimeric oligonucleotides it was found that whereas the enzymes can bind both DNA and RNA, they could translocate only along DNA and only DNA stimulates the ATPase activity of the enzymes. Recent observations suggest that helicases may interact with enzymes participating in RNA metabolism and that RNA-DNA hybrids may be present on the chromosomes. Thus, the results presented here may suggest a new role for the replicative helicases during chromosomal replication or in other cellular processes.
Shin
Jae-Ho
JH
University of Maryland Biotechnology Institute, Center for Advanced Research in Biotechnology, Rockville, Maryland 20850, USA.
Kelman
Zvi
Z
D000251:Adenosine Triphosphatases / Q000737:chemistry; D019843:Archaeal Proteins / Q000378:metabolism; D019921:Biotinylation; D018797:Cell Cycle Proteins / Q000737:chemistry; D004247:DNA / Q000737:chemistry*; D004265:DNA Helicases / Q000737:chemistry* / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D017014:Methanobacterium / Q000378:metabolism; D064151:Minichromosome Maintenance Complex Component 6; D009691:Nucleic Acid Denaturation; D012313:RNA / Q000737:chemistry*; D012568:Schizosaccharomyces / Q000378:metabolism; D029702:Schizosaccharomyces pombe Proteins / Q000737:chemistry; D019809:Streptavidin / Q000737:chemistry
D019843:Archaeal Proteins; D018797:Cell Cycle Proteins; D029702:Schizosaccharomyces pombe Proteins; D012313:RNA; D004247:DNA; D019809:Streptavidin; D000251:Adenosine Triphosphatases; D004265:DNA Helicases; D064151:Minichromosome Maintenance Complex Component 6; C092300:mcm6 protein, S pombe; C403162:MCM protein, Methanobacterium thermoautotrophicum