title: Deamination intensity profiling of human APOBEC3 protein activity along the near full-length genomes of HIV-1 and MoMLV by HyperHRM analysis.
abstract: Enzymatic deamination of cytidines in DNA is an intrinsic component of antibody maturation and retroviral resistance, but can also be a source of HIV drug resistance and cancer-causing mutations. Here, we developed a high-throughput method based on high resolution melt (HRM) analysis called HyperHRM that can screen genomic DNA for rare hypermutated proviral sequences and accurately quantify the number of C-to-T or G-to-A mutations in each sequence. We demonstrate the effectiveness of the approach by profiling in parallel the intensity of the DNA mutator activity of all seven human APOBEC3 proteins on the near full-length sequence of HIV-1 and the Moloney murine leukemia virus. Additionally, HRM was successfully used to identify hypermutated proviral sequences in peripheral blood mononuclear cells from an HIV-1 patient. These results exemplify the effectiveness of HRM-based approaches for hypermutation quantification and for the detection of hypermutated DNA sequences potentially associated with disease or retroviral drug resistance.
authors: Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of Ottawa, Ottawa, Ontario, Canada.
authors: Kasandra
authors: K
authors: Bélanger
authors: Mathieu
authors: M
authors: Savoie
authors: Halil
authors: H
authors: Aydin
authors: Tyler Milston
authors: TM
authors: Renner
authors: Zahra
authors: Z
authors: Montazeri
authors: Marc-André
authors: MA
authors: Langlois
mesh_terms: D000071478:APOBEC Deaminases; D000818:Animals; D002460:Cell Line; D019295:Computational Biology; D003564:Cytidine Deaminase; D043525:Cytosine Deaminase / Q000378:metabolism*; D004252:DNA Mutational Analysis / Q000295:instrumentation / Q000379:methods*; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics; D003641:Deamination; D016679:Genome, Viral; D015658:HIV Infections / Q000201:enzymology* / Q000821:virology; D015497:HIV-1 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000821:virology; D051379:Mice; D008979:Moloney murine leukemia virus / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D009691:Nucleic Acid Denaturation; D017354:Point Mutation; D012192:Retroviridae Infections / Q000201:enzymology* / Q000821:virology
chemical_list: D004279:DNA, Viral; D043525:Cytosine Deaminase; D000071478:APOBEC Deaminases; C502889:APOBEC3 proteins, human; D003564:Cytidine Deaminase
keywords: APOBEC3; Cytidine deamination; HIV; Hypermutation; Moloney murine leukemia virus