Bromodeoxyuridine

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Bromodeoxyuridine
Bromodeoxyuridine.svg
Names
IUPAC name
5-Bromo-2′-deoxyuridine
Systematic IUPAC name
5-Bromo-1-[(2R,4S,5R)-4-hydroxy-5-(hydroxymethyl)oxolan-2-yl]pyrimidine-2,4(1H,3H)-dione
Identifiers
3D model (JSmol)
ChEMBL
ChemSpider
ECHA InfoCard 100.000.378 OOjs UI icon edit-ltr-progressive.svg
MeSH Bromodeoxyuridine
PubChem CID
UNII
  • InChI=1S/C9H11BrN2O5/c10-4-2-12(9(16)11-8(4)15)7-1-5(14)6(3-13)17-7/h2,5-7,13-14H,1,3H2,(H,11,15,16)/t5-,6+,7+/m0/s1 X mark.svgN
    Key: WOVKYSAHUYNSMH-RRKCRQDMSA-N X mark.svgN
  • InChI=1/C9H11BrN2O5/c10-4-2-12(9(16)11-8(4)15)7-1-5(14)6(3-13)17-7/h2,5-7,13-14H,1,3H2,(H,11,15,16)/t5-,6+,7+/m0/s1
    Key: WOVKYSAHUYNSMH-RRKCRQDMBM
  • c1c(c(=O)[nH]c(=O)n1[C@H]2C[C@@H]([C@H](O2)CO)O)Br
Properties
C9H11BrN2O5
Molar mass 307.100 g·mol−1
Except where otherwise noted, data are given for materials in their standard state (at 25 °C [77 °F], 100 kPa).
X mark.svgN  verify  (what is  Yes check.svgYX mark.svgN ?)

Bromodeoxyuridine (5-bromo-2'-deoxyuridine, BrdU, BUdR, BrdUrd, broxuridine) is a synthetic nucleoside analogue with a chemical structure similar to thymidine. BrdU is commonly used to study cell proliferation in living tissues [1] and has been studied as a radiosensitizer [2] and diagnostic tool in people with cancer. [3]

Contents

During the S phase of the cell cycle (when DNA replication occurs), BrdU can be incorporated in place of thymidine in newly synthesized DNA molecules of dividing cells. [4] Cells that have recently performed DNA replication or DNA repair can be detected with antibodies specific for BrdU using techniques such as immunohistochemistry or immunofluorescence. [5] BrdU-labelled cells in humans can be detected up to two years after BrdU infusion. [6]

Because BrdU can replace thymidine during DNA replication, it can cause mutations, and its use is therefore potentially a health hazard.[ citation needed ] However, because it is neither radioactive nor myelotoxic at labeling concentrations, it is widely preferred for in vivo studies of cancer cell proliferation. [7] [8] However, at radiosensitizing concentrations, BrdU becomes myelosuppressive, thus limiting its use for radiosensitizing. [2]

BrdU differs from thymidine in that BrdU substitutes a bromine atom for thymidine's CH3 group. The Br substitution can be used in X-ray diffraction experiments in crystals containing either DNA or RNA. The Br atom acts as an anomalous scatterer and its larger size will affect the crystal's X-ray diffraction enough to detect isomorphous differences as well. [9] [10]

Bromodeoxyuridine releases gene silencing caused by DNA methylation. [11]

BrdU can also be used to identify microorganisms that respond to specific carbon substrates in aquatic [12] and soil [13] environments. A carbon substrate added to the incubations of environmental samples will cause the growth of microorganisms that can utilize that substrate. These microorganisms will then incorporate BrdU into their DNA as they grow. Community DNA can then be isolated and BrdU-labeled DNA purified using an immunocapture technique. [14] Subsequent sequencing of the labeled DNA can then be used to identify the microbial taxa that participated in the degradation of the added carbon source.

However, it is not certain whether all microbes present in an environmental sample can incorporate BrdU into their biomass during de novo DNA synthesis. Therefore, a group of microorganisms may respond to a carbon source but go undetected using this technique. Additionally, this technique is biased towards identifying microorganisms with A- and T-rich genomes.

DNA with BrdU transcribes as usual DNA, with guanine included in RNA as a complement to BrdU. [15]

See also

Related Research Articles

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Thymidine, also known as deoxythymidine, deoxyribosylthymine, or thymine deoxyriboside, is a pyrimidine deoxynucleoside. Deoxythymidine is the DNA nucleoside T, which pairs with deoxyadenosine (A) in double-stranded DNA. In cell biology it is used to synchronize the cells in G1/early S phase. The prefix deoxy- is often left out since there are no precursors of thymine nucleotides involved in RNA synthesis.

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Thymidine kinase is an enzyme, a phosphotransferase : 2'-deoxythymidine kinase, ATP-thymidine 5'-phosphotransferase, EC 2.7.1.21. It can be found in most living cells. It is present in two forms in mammalian cells, TK1 and TK2. Certain viruses also have genetic information for expression of viral thymidine kinases. Thymidine kinase catalyzes the reaction:

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<span class="mw-page-title-main">Nucleoside analogue</span> Biochemical compound

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HAT Medium is a selection medium for mammalian cell culture, which relies on the combination of aminopterin, a drug that acts as a powerful folate metabolism inhibitor by inhibiting dihydrofolate reductase, with hypoxanthine and thymidine which are intermediates in DNA synthesis. The trick is that aminopterin blocks DNA de novo synthesis, which is absolutely required for cell division to proceed, but hypoxanthine and thymidine provide cells with the raw material to evade the blockage, provided that they have the right enzymes, which means having functioning copies of the genes that encode them.

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<span class="mw-page-title-main">Thymidylate synthase</span> Enzyme

Thymidylate synthase (TS) is an enzyme that catalyzes the conversion of deoxyuridine monophosphate (dUMP) to deoxythymidine monophosphate (dTMP). Thymidine is one of the nucleotides in DNA. With inhibition of TS, an imbalance of deoxynucleotides and increased levels of dUMP arise. Both cause DNA damage.

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5-Ethynyl-2′-deoxyuridine (EdU) is a thymidine analogue which is incorporated into the DNA of dividing cells. EdU is used to assay DNA synthesis in cell culture and detect cells in embryonic, neonatal and adult animals which have undergone DNA synthesis. Whilst at high doses it can be cytotoxic, this molecule is now widely used to track proliferating cells in multiple biological systems.

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References

  1. ↑ Lehner, Bernadette; Sandner, Beatrice; Marschallinger, Julia; Lehner, Christine; Furtner, Tanja; Couillard-Despres, Sebastien; Rivera, Francisco J.; Brockhoff, Gero; Bauer, Hans-Christian; Weidner, Norbert; Aigner, Ludwig (2011). "The dark side of BrdU in neural stem cell biology: Detrimental effects on cell cycle, differentiation and survival". Cell and Tissue Research. 345 (3): 313–28. doi:10.1007/s00441-011-1213-7. PMID   21837406. S2CID   756261.
  2. 1 2 Russo A, Gianni L, Kinsella TJ, Klecker RW, Jenkins J, Rowland J, Glatstein E, Mitchell JB, Collins J, Myers C (1984). "Pharmacological evaluation of intravenous delivery of 5-bromodeoxyuridine to patients with brain tumors". Cancer Res. 44 (4): 1702–5. PMID   6704976.
  3. ↑ Dolbeare, F (May 1995). "Bromodeoxyuridine: a diagnostic tool in biology and medicine, Part I: Historical perspectives, histochemical methods and cell kinetics". The Histochemical Journal. 27 (5): 339–69. doi:10.1007/BF02389022. PMID   7657555. S2CID   21785471.
  4. ↑ Kee, N; S Sivalingam; R Boonstra; J.M Wojtowicz (March 2002). "The utility of Ki-67 and BrdU as proliferative markers of adult neurogenesis". Journal of Neuroscience Methods. 115 (1): 97–105. doi:10.1016/S0165-0270(02)00007-9. hdl: 1807/357 . PMID   11897369. S2CID   17572667.
  5. ↑ Konishi, Teruaki; Takeyasu, Akihiro; Natsume, Toshiyuki; Furusawa, Yoshiya; Hieda, Kotaro (2011). "Visualization of Heavy Ion Tracks by Labeling 3'-OH Termini of Induced DNA Strand Breaks". Journal of Radiation Research. 52 (4): 433–40. Bibcode:2011JRadR..52..433K. doi:10.1269/jrr.10097. PMID   21785232.
  6. ↑ Eriksson, Peter; Ekaterina Perfilieva; Thomas Björk-Eriksson; Ann-Marie Alborn; Claes Nordborg; Daniel A. Peterson; Fred H. Gage (1998). "Neurogenesis in the adult human hippocampus". Nature Medicine. 1313-1317. 4 (11): 1313–1317. doi: 10.1038/3305 . PMID   9809557.
  7. ↑ Fujimaki, Takamitsu; Masao Matsutani; Osamu Nakamura; Akio Asai; Nobuaki Funada; Morio Koike; Hiromu Segawa; Kouichi Aritake; Takanori Fukushima; Shuntaro Houjo; Akira Tamura; Keiji Sano (29 June 2006). "Correlation Between Bromodeoxyuridine- Labeling Indices and Patient Prognosis in Cerebral Astrocytic Tumors of Adults". Cancer. 67 (6): 1629–1634. doi: 10.1002/1097-0142(19910315)67:6<1629::AID-CNCR2820670626>3.0.CO;2-E . PMID   2001552.
  8. ↑ Hoshino, Takao; Tadashi Nagashima; Judith Murovic; Ellen M. Levin; Victor A. Levin; Stephen M. Rupp (1985). "Cell Kinetic Studies of In Situ Human Brain Tumors With Bromodeoxyuridine". Cytometry. 6 (6): 627–632. doi: 10.1002/cyto.990060619 . PMID   2998714.
  9. ↑ Peterson, M. R.; Harrop, S. J.; McSweeney, S. M.; Leonard, G. A.; Thompson, A. W.; Hunter, W. N.; Helliwell, J. R. (1996). "MAD Phasing Strategies Explored with a Brominated Oligonucleotide Crystal at 1.65Å Resolution". Journal of Synchrotron Radiation. 3 (Pt 1): 24–34. Bibcode:1996JSynR...3...24P. doi: 10.1107/S0909049595013288 . PMID   16702655.
  10. ↑ Beck, Tobias; Gruene, Tim; Sheldrick, George M. (2010). "The magic triangle goes MAD: Experimental phasing with a bromine derivative". Acta Crystallographica Section D. 66 (4): 374–80. Bibcode:2010AcCrD..66..374B. doi:10.1107/S0907444909051609. PMC   2852301 . PMID   20382990.
  11. ↑ Weiss RA (2013). "On the concept and elucidation of endogenous retroviruses". Philos. Trans. R. Soc. Lond. B Biol. Sci. 368 (1626): 20120494. doi:10.1098/rstb.2012.0494. PMC   3758183 . PMID   23938748.
  12. ↑ Tada, Yuya; Grossart, Hans-Peter (2013). "Community shifts of actively growing lake bacteria after N-acetyl-glucosamine addition: improving the BrdU-FACS method". The ISME Journal. 8 (2): 441–454. doi:10.1038/ismej.2013.148. ISSN   1751-7362. PMC   3906810 . PMID   23985742.
  13. ↑ Borneman J (1999). "Culture-independent identification of microorganisms that respond to specified stimuli". Appl. Environ. Microbiol. 65 (8): 3398–400. Bibcode:1999ApEnM..65.3398B. doi:10.1128/AEM.65.8.3398-3400.1999. PMC   91510 . PMID   10427025.
  14. ↑ Urbach, Ena; Kevin L. Vergin; Stephen J. Giovannoni (March 1999). "Immunochemical Detection and Isolation of DNA from Metabolically Active Bacteria". Applied and Environmental Microbiology. 65 (3): 1207–1213. Bibcode:1999ApEnM..65.1207U. doi:10.1128/AEM.65.3.1207-1213.1999. PMC   91166 . PMID   10049885.
  15. ↑ Hill BT, Baserga R (1975). "Effect of 5-bromodeoxyuridine on the transcriptional properties of the genome in WI-38 human diploid fibroblasts". Chem. Biol. Interact. 10 (5): 363–375. Bibcode:1975CBI....10..363H. doi:10.1016/0009-2797(75)90058-7. PMID   1095238.
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