Transient expression and activity of human DNA polymerase iota in loach embryos

• Published in: Biotechnology letters 2012-02, Vol.34 (2), p.205-212
• Main Authors: Makarova, Irina V, Kazakov, Andrey A, Makarova, Alena V, Khaidarova, Nella V, Kozikova, Larisa V, Nenasheva, Valentina V, Gening, Leonid V, Tarantul, Vyacheslav Z, Andreeva, Ludmila E
• Format: Article
• Language: English
• Subjects: abnormal development; Animal models; Animals; Applied Microbiology; Biochemistry; Biomedical and Life Sciences; Biotechnology; Cell Extracts; Cell Nucleus - pathology; Congenital Abnormalities - pathology; Cypriniformes - genetics; Deoxyribonucleic acid; DNA; DNA polymerase; DNA-directed DNA polymerase; DNA-Directed DNA Polymerase - metabolism; Embryo, Nonmammalian; Embryos; fish; Gene Expression; Humans; Life Sciences; Microbiology; Misgurnus; Misgurnus fossilis; neoplasms; Original Research Paper; Sensitivity and Specificity; thymine
• ISSN: 0141-5492; 1573-6776
• DOI: 10.1007/s10529-011-0764-8

Human DNA polymerase iota (Pol ι) is a Y-family DNA polymerase with unusual biochemical properties and not fully understood functions. Pol ι preferentially incorporates dGTP opposite template thymine. This property can be used to monitor Pol ι activity in the presence of other DNA polymerases, e.g. in cell extracts of tissues and tumors. We have now confirmed the specificity and sensitivity of the method of Pol ι activity detection in cell extracts using an animal model of loach Misgurnus fossilis embryos transiently expressing human Pol ι. The overexpression of Pol ι was shown to be accompanied by an increase in abnormalities in development and the frequency of pycnotic nuclei in fish embryos. Further analysis of fish embryos with constitutive or regulated Pol ι expression may provide insights into Pol ι functions in vertebrate animals.