title: Human apurinic/apyrimidinic endonuclease 1 (APE1) has 3' RNA phosphatase and 3' exoribonuclease activities.
abstract: Apurinic/apyrimidinic endonuclease 1 (APE1) is the predominant mammalian enzyme in DNA base excision repair pathway that cleaves the DNA backbone immediately 5' to abasic sites. In addition to its abasic endonuclease activity, APE1 has 3' phosphatase and 3'-5' exonuclease activities against DNA. We recently identified APE1 as an endoribonuclease that preferentially cleaves at UA, UG, and CA sites in single-stranded regions of RNAs and can regulate c-myc mRNA level and half-life in cells. APE1 can also endonucleolytically cleave abasic single-stranded RNA. Here, we show for the first time that the human APE1 has 3' RNA phosphatase and 3' exoribonuclease activities. Using three distinct RNA substrates, we show that APE1, but not RNase A, can remove the phosphoryl group from the 3' end of RNA decay products. Studies using various site-directed APE1 mutant proteins (H309N, H309S, D283N, N68A, D210N, Y171F, D308A, F266A, and D70A) suggest that the 3' RNA phosphatase activity shares the same active center as its other known nuclease activities. A number of APE1 variants previously identified in the human population, including the most common D148E variant, have greater than 80% reduction in the 3' RNA phosphatase activity. APE1 can remove a ribonucleotide from the 3' overhang of RNA decay product, but its 3'-5' exoribonuclease activity against unstructured poly(A), poly(C), and poly(U) RNAs is relatively weak. This study further underscores the significance of understanding the role of APE1 in RNA metabolism in vivo.
authors.affiliation: Chemistry Program, University of Northern British Columbia, 3333 University Way, Prince George, British Columbia, V2N 4Z9, Canada.
authors.forename: Manbir
authors.initials: M
authors.lastname: Chohan
authors.affiliation: Chemistry Program, University of Northern British Columbia, 3333 University Way, Prince George, British Columbia, V2N 4Z9, Canada.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Mackedenski
authors.affiliation: Chemistry Program, University of Northern British Columbia, 3333 University Way, Prince George, British Columbia, V2N 4Z9, Canada.
authors.forename: Wai-Ming
authors.initials: WM
authors.lastname: Li
authors.affiliation: Chemistry Program, University of Northern British Columbia, 3333 University Way, Prince George, British Columbia, V2N 4Z9, Canada. Electronic address: chow.lee@unbc.ca.
authors.forename: Chow H
authors.initials: CH
authors.lastname: Lee
mesh_terms: D016227:Benzoquinones / Q000494:pharmacology; D004260:DNA Repair / Q000187:drug effects; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase / Q000235:genetics / Q000378:metabolism*; D004722:Endoribonucleases / Q000235:genetics / Q000378:metabolism; D005095:Exoribonucleases / Q000235:genetics / Q000378:metabolism*; D005419:Flavonoids / Q000494:pharmacology; D006207:Half-Life; D006801:Humans; D006898:Hydroxylamines / Q000494:pharmacology; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics / Q000378:metabolism*; D011422:Propionates / Q000494:pharmacology; D012313:RNA / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D016227:Benzoquinones; D005419:Flavonoids; D006898:Hydroxylamines; D011422:Propionates; C075569:E 3330; D012313:RNA; C040015:myricetin; C005214:methoxyamine; D004722:Endoribonucleases; D005095:Exoribonucleases; D010744:Phosphoric Monoester Hydrolases; C085108:APEX1 protein, human; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase
keywords: 3′ RNA phosphatase; APE1; RNA