Terminal deoxynucleotidyl transferase
Terminal deoxynucleotidyl transferase (TDT), also known as DNA nucleotidylexotransferase (DNTT) or terminal transferase, is a specialized DNA polymerase expressed in immature, pre-B, pre-T lymphoid cells, and acute lymphoblastic leukemia/lymphoma cells. TdT adds N-nucleotides to the V,D, and J exons during antibody gene recombination enabling the phenomenon of junctional diversity. In humans, terminal transferase is encoded by the DNTT gene.[1][2]
TdT is notably absent in fetal liver HSCs significantly impairing junctional diversity in B-cells during the fetal period.[3]
Function
TdT catalyses the addition of nucleotides to the 3' terminus of a DNA molecule. Unlike most DNA polymerases it does not require a template. The preferred substrate of this enzyme is a 3'-overhang, but it can also add nucleotides to blunt or recessed 3' ends. Cobalt is a necessary cofactor, however the enzyme catalyzes reaction upon Mg and Mn administration in vitro.
Uses
Terminal transferase has applications in molecular biology. It can be used in RACE to add nucleotides which can then be used as a template for a primer in subsequent PCR. It can also be used to add nucleotides labeled with radioactive isotopes, for example in the TUNEL assay (Terminal deoxynucleotidyl transferase dUTP Nick End Labeling) for the demonstration of apoptosis (which is marked, in part, by fragmented DNA). Also used in the immunofluorescence assay for the diagnosis of acute lymphoblastic leukemia.[4]
In immunohistochemistry, antibodies to TdT can be used to demonstrate the presence of immature T and B cells and multipotent haematopoietic stem cells, which possess the antigen, while mature lymphoid cells are always TdT-negative. While TdT-positive cells are found in small numbers in healthy lymph nodes and tonsils, the malignant cells of acute lymphoblastic leukaemia are also TdT positive, and the antibody can therefore be used as part of a panel to diagnose this disease and to distinguish it from, for example, small cell tumours of childhood.[5]
See also
References
- ^ Isobe M, Huebner K, Erikson J, Peterson RC, Bollum FJ, Chang LM, Croce CM (September 1985). "Chromosome localization of the gene for human terminal deoxynucleotidyltransferase to region 10q23-q25". Proc. Natl. Acad. Sci. U.S.A. 82 (17): 5836–40. doi:10.1073/pnas.82.17.5836. PMC 390648. PMID 3862101. http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pmcentrez&artid=390648.
- ^ Yang-Feng TL, Landau NR, Baltimore D, Francke U (1986). "The terminal deoxynucleotidyltransferase gene is located on human chromosome 10 (10q23-q24) and on mouse chromosome 19". Cytogenet. Cell Genet. 43 (3-4): 121–6. doi:10.1159/000132309. PMID 3467897.
- ^ Hardy, Richard (2008). "Chapter 7: B Lymphocyte Development and Biology". In Paul, William (Book). Fundamental Immunology (6th ed.). Philidelphia, PA: Lippincott Williams & Wilkins. pp. 237–269. ISBN 0-7817-6519-6.
- ^ Faber J, Kantarjian H, Roberts MW, Keating M, Freireich E, Albitar M (January 2000). "Terminal deoxynucleotidyl transferase-negative acute lymphoblastic leukemia". Arch. Pathol. Lab. Med. 124 (1): 92–7. PMID 10629138.
- ^ Leong, Anthony S-Y; Cooper, Kumarason; Leong, F Joel W-M (2003). Manual of Diagnostic Cytology (2 ed.). Greenwich Medical Media, Ltd.. pp. 413-414. ISBN 1-84110-100-1.
Further reading
- O'Malley DP, Orazi A (2006). "Terminal deoxynucleotidyl transferase-positive cells in spleen, appendix and branchial cleft cysts in pediatric patients.". Haematologica 91 (8): 1139–40. PMID 16885057.
- Yamashita N, Shimazaki N, Ibe S, et al. (2001). "Terminal deoxynucleotidyltransferase directly interacts with a novel nuclear protein that is homologous to p65.". Genes Cells 6 (7): 641–52. doi:10.1046/j.1365-2443.2001.00449.x. PMID 11473582.
- Chang LM, Bollum FJ (1986). "Molecular biology of terminal transferase.". CRC Crit. Rev. Biochem. 21 (1): 27–52. doi:10.3109/10409238609113608. PMID 3524991.
- Maezawa S, Hayano T, Koiwai K, et al. (2008). "Bood POZ containing gene type 2 is a human counterpart of yeast Btb3p and promotes the degradation of terminal deoxynucleotidyltransferase.". Genes Cells 13 (5): 439–57. doi:10.1111/j.1365-2443.2008.01179.x. PMID 18429817.
- Taplin ME, Frantz ME, Canning C, et al. (1996). "Evidence against T-cell development in the adult human intestinal mucosa based upon lack of terminal deoxynucleotidyltransferase expression.". Immunology 87 (3): 402–7. doi:10.1046/j.1365-2567.1996.496571.x. PMC 1384108. PMID 8778025. http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pmcentrez&artid=1384108.
- Grupe A, Li Y, Rowland C, et al. (2006). "A scan of chromosome 10 identifies a novel locus showing strong association with late-onset Alzheimer disease.". Am. J. Hum. Genet. 78 (1): 78–88. doi:10.1086/498851. PMC 1380225. PMID 16385451. http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pmcentrez&artid=1380225.
- Dworzak MN, Fritsch G, Fröschl G, et al. (1998). "Four-color flow cytometric investigation of terminal deoxynucleotidyl transferase-positive lymphoid precursors in pediatric bone marrow: CD79a expression precedes CD19 in early B-cell ontogeny.". Blood 92 (9): 3203–9. PMID 9787156.
- Fujita K, Shimazaki N, Ohta Y, et al. (2003). "Terminal deoxynucleotidyltransferase forms a ternary complex with a novel chromatin remodeling protein with 82 kDa and core histone.". Genes Cells 8 (6): 559–71. doi:10.1046/j.1365-2443.2003.00656.x. PMID 12786946.
- Kubota T, Maezawa S, Koiwai K, et al. (2007). "Identification of functional domains in TdIF1 and its inhibitory mechanism for TdT activity.". Genes Cells 12 (8): 941–59. doi:10.1111/j.1365-2443.2007.01105.x. PMID 17663723.
- Suzuki Y, Yoshitomo-Nakagawa K, Maruyama K, et al. (1997). "Construction and characterization of a full length-enriched and a 5'-end-enriched cDNA library.". Gene 200 (1-2): 149–56. doi:10.1016/S0378-1119(97)00411-3. PMID 9373149.
- Bridges SL (1998). "Frequent N addition and clonal relatedness among immunoglobulin lambda light chains expressed in rheumatoid arthritis synovia and PBL, and the influence of V lambda gene segment utilization on CDR3 length.". Mol. Med. 4 (8): 525–53. PMC 2230400. PMID 9742508. http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pmcentrez&artid=2230400.
- Liu L, McGavran L, Lovell MA, et al. (2004). "Nonpositive terminal deoxynucleotidyl transferase in pediatric precursor B-lymphoblastic leukemia.". Am. J. Clin. Pathol. 121 (6): 810–5. doi:10.1309/QD18-PPV1-NH3T-EUTF. PMID 15198352.
- Yang B, Gathy KN, Coleman MS (1994). "Mutational analysis of residues in the nucleotide binding domain of human terminal deoxynucleotidyl transferase.". J. Biol. Chem. 269 (16): 11859–68. PMID 8163485.
- Gerhard DS, Wagner L, Feingold EA, et al. (2004). "The status, quality, and expansion of the NIH full-length cDNA project: the Mammalian Gene Collection (MGC).". Genome Res. 14 (10B): 2121–7. doi:10.1101/gr.2596504. PMC 528928. PMID 15489334. http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pmcentrez&artid=528928.
- Thai TH, Kearney JF (2004). "Distinct and opposite activities of human terminal deoxynucleotidyltransferase splice variants.". J. Immunol. 173 (6): 4009–19. PMID 15356150.
- Shimazaki N, Fujita K, Koiwai O (2002). "[Expression and function of terminal deoxynucleotidyl-transferase and discovery of novel DNA polymerase mu]". Seikagaku 74 (3): 227–32. PMID 11974916.
- Mahajan KN, Mitchell BS (2003). "Role of human Pso4 in mammalian DNA repair and association with terminal deoxynucleotidyl transferase.". Proc. Natl. Acad. Sci. U.S.A. 100 (19): 10746–51. doi:10.1073/pnas.1631060100. PMC 196874. PMID 12960389. http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pmcentrez&artid=196874.
- Strausberg RL, Feingold EA, Grouse LH, et al. (2002). "Generation and initial analysis of more than 15,000 full-length human and mouse cDNA sequences.". Proc. Natl. Acad. Sci. U.S.A. 99 (26): 16899–903. doi:10.1073/pnas.242603899. PMC 139241. PMID 12477932. http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pmcentrez&artid=139241.
- Mahajan KN, Gangi-Peterson L, Sorscher DH, et al. (1999). "Association of terminal deoxynucleotidyl transferase with Ku.". Proc. Natl. Acad. Sci. U.S.A. 96 (24): 13926–31. doi:10.1073/pnas.96.24.13926. PMC 24167. PMID 10570175. http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pmcentrez&artid=24167.
- Ibe S, Fujita K, Toyomoto T, et al. (2001). "Terminal deoxynucleotidyltransferase is negatively regulated by direct interaction with proliferating cell nuclear antigen.". Genes Cells 6 (9): 815–24. doi:10.1046/j.1365-2443.2001.00460.x. PMID 11554927.
External links