Base excision repair

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Basic steps of base excision repair

In biochemistry and genetics, base excision repair (BER) is a cellular mechanism that repairs damaged DNA throughout the cell cycle. It is responsible primarily for removing small, non-helix-distorting base lesions from the genome. The related nucleotide excision repair pathway repairs bulky helix-distorting lesions. BER is important for removing damaged bases that could otherwise cause mutations by mispairing or lead to breaks in DNA during replication. BER is initiated by DNA glycosylases, which recognize and remove specific damaged or inappropriate bases, forming AP sites. These are then cleaved by an AP endonuclease. The resulting single-strand break can then be processed by either short-patch (where a single nucleotide is replaced) or long-patch BER (where 2-10 new nucleotides are synthesized).[1]

Lesions processed by BER[edit]

8-oxoguanine forms a Hoogsteen base pair with adenine

Single bases in DNA can be chemically damaged by a variety of mechanisms, the most common ones being deamination, oxidation, and alkylation. These modifications can affect the ability of the base to hydrogen-bond, resulting in incorrect base-pairing, and, as a consequence, mutations in the DNA. For example, incorporation of adenine across from 8-oxoguanine (right) during DNA replication causes a G:C base pair to be mutated to T:A. Other examples of base lesions repaired by BER include:

In addition to base lesions, the downstream steps of BER are also utilized to repair single-strand breaks.

The choice between long-patch and short-patch repair[edit]

The choice between short- and long-patch repair is currently under investigation. Various factors are thought to influence this decision, including the type of lesion, the cell cycle stage, and whether the cell is terminally differentiated or actively dividing.[3] Some lesions, such as oxidized or reduced AP sites, are resistant to pol β lyase activity and, therefore, must be processed by long-patch BER.

Pathway preference may differ between organisms, as well. While human cells utilize both short- and long-patch BER, the yeast Saccharomyces cerevisiae was long thought to lack a short-patch pathway because it does not have homologs of several mammalian short-patch proteins, including pol β, DNA ligase III, XRCC1, and the kinase domain of PNKP. The recent discovery that the poly-A polymerase Trf4 possesses 5' dRP lyase activity has challenged this view.[4]

Proteins involved in base excision repair[edit]

DNA glycosylases[edit]

Main article: DNA glycosylase
Uracil DNA glycosylase flips a uracil residue out of the duplex, shown in yellow.

DNA glycosylases are responsible for initial recognition of the lesion. They flip the damaged base out of the double helix, as pictured, and cleave the N-glycosidic bond of the damaged base, leaving an AP site. There are two categories of glycosylases: monofunctional and bifunctional. Monofunctional glycosylases have only glycosylase activity, whereas bifunctional glycosylases also possess AP lyase activity. Therefore, bifunctional glycosylases can convert a base lesion into a single-strand break without the need for an AP endonuclease. β-Elimination of an AP site by a glycosylase-lyase yields a 3' α,β-unsaturated aldehyde adjacent to a 5' phosphate, which differs from the AP endonuclease cleavage product.[5] Some glycosylase-lyases can further perform δ-elimination, which converts the 3' aldehyde to a 3' phosphate. A wide variety of glycosylases have evolved to recognize different damaged bases. Examples of DNA glycosylases include Ogg1, which recognizes 8-oxoguanine, Mag1, which recognizes 3-methyladenine, and UNG, which removes uracil from DNA.

AP endonucleases[edit]

Main article: AP endonuclease

The AP endonucleases cleave an AP site to yield a 3' hydroxyl adjacent to a 5' deoxyribosephosphate (dRP). AP endonucleases are divided into two families based on their homology to the ancestral bacterial AP endonucleaes endonuclease IV and exonuclease III.[6] Many eukaryotes have members of both families, including the yeast Saccharomyces cerevisiae, in which Apn1 is the EndoIV homolog and Apn2 is related to ExoIII. In humans, two AP endonucleases, APEX1 and APEX2, have been identified.[7] It is a member of the ExoIII family.

End processing enzymes[edit]

In order for ligation to occur, a DNA strand break must have a hydroxyl on its 3' end and a phosphate on its 5' end. In humans, polynucleotide kinase-phosphatase (PNKP) promotes formation of these ends during BER. This protein has a kinase domain, which phosphorylates 5' hydroxyl ends, and a phosphatase domain, which removes phosphates from 3' ends. Together, these activities ready single-strand breaks with damaged termini for ligation. The AP endonucleases also participate in 3' end processing. Besides opening AP sites, they possess 3' phosphodiesterase activity and can remove a variety of 3' lesions including phosphates, phosphoglycolates, and aldehydes. 3'-Processing must occur before DNA synthesis can initiate because DNA polymerases require a 3' hydroxyl to extend from.

DNA polymerases[edit]

Main article: DNA Polymerase

Pol β is the main human polymerase that catalyzes short-patch BER, with pol λ able to compensate in its absence.[8] These polymerases are members of the Pol X family and typically insert only a single nucleotide. In addition to polymerase activity, these enzymes have a lyase domain that removes the 5' dRP left behind by AP endonuclease cleavage. During long-patch BER, DNA synthesis is thought to be mediated by pol δ and pol ε along with the processivity factor PCNA, the same polymerases that carry out DNA replication. These polymerases perform displacing synthesis, meaning that the downstream 5' DNA end is "displaced" to form a flap (see diagram above). Pol β can also perform long-patch displacing synthesis and can, therefore, participate in either BER pathway.[9] Long-patch synthesis typically inserts 2-10 new nucleotides.

Flap endonuclease[edit]

Main article: Flap endonuclease

FEN1 removes the 5' flap generated during long patch BER. This endonuclease shows a strong preference for a long 5' flap adjacent to a 1-nt 3' flap.[10] The yeast homolog of FEN1 is RAD27. In addition to its role in long-patch BER, FEN1 cleaves flaps with a similar structure during Okazaki fragment processing, an important step in lagging strand DNA replication.

DNA ligase[edit]

Main article: DNA ligase

DNA ligase III along with its cofactor XRCC1 catalyzes the nick-sealing step in short-patch BER in humans. DNA ligase I ligates the break in long-patch BER.

Links between BER and cancer[edit]

Defects in a variety of DNA repair pathways lead to cancer predisposition, and BER appears to follow this pattern. Deletion mutations in BER genes have shown to result in a higher mutation rate in a variety of organisms, implying that loss of BER could contribute to the development of cancer. Indeed, somatic mutations in Pol β have been found in 30% of human cancers, and some of these mutations lead to transformation when expressed in mouse cells.[11] Mutations in the DNA glycosylase MYH are also known to increase susceptibility to colon cancer.[12]

See also[edit]


  1. ^ Liu Y, Prasad R, Beard WA, Kedar PS, Hou EW, Shock DD, Wilson SH, (2007). "Coordination of Steps in Single-nucleotide Base Excision Repair Mediated by Apurinic/Apyrimidinic Endonuclease 1 and DNA Polymerase β". Journal of Biological Chemistry 282 (18): 13532–13541. doi:10.1074/jbc.M611295200. PMC 2366199. PMID 17355977. 
  2. ^ Jayanta Chaudhuri & Frederick W. Alt (2004). "Class-switch recombination: interplay of transcription, DNA deamination and DNA repair". Nature Reviews Immunology 4 (7): 541–552. doi:10.1038/nri1395. PMID 15229473. 
  3. ^ Fortini P, Dogliotti E (April 2007). "Base damage and single-strand break repair: mechanisms and functional significance of short- and long-patch repair subpathways". DNA Repair 6 (4): 398–409. doi:10.1016/j.dnarep.2006.10.008. PMID 17129767. 
  4. ^ Gellon L, Carson DR, Carson JP, Demple B (February 2008). "Intrinsic 5'-Deoxyribose-5-phosphate Lyase Activity in Saccharomyces cerevisiae Trf4 Protein with a Possible Role in Base Excision DNA Repair". DNA Repair 7 (2): 187–98. doi:10.1016/j.dnarep.2007.09.009. PMC 2258243. PMID 17983848. 
  5. ^ Fromme JC, Banerjee A, Verdine GL (February 2004). "DNA glycosylase recognition and catalysis". Curr. Opin. Struct. Biol. 14 (1): 43–9. doi:10.1016/ PMID 15102448. 
  6. ^ Aravind, L., Walker, D.R. & Koonin, E.V. (1999). "Conserved domains in DNA repair proteins and evolution of repair systems". Nucleic Acids Research 27 (5): 1223–1242. doi:10.1093/nar/27.5.1223. PMC 148307. PMID 9973609. 
  7. ^ Demple, B., Herman, T. & Chen, D.S. (1991). "Cloning and expression of APE, the cDNA encoding the major human apurinic endonuclease: definition of a family of DNA repair enzymes". Pnas USA 88 (24): 11450–11454. doi:10.1073/pnas.88.24.11450. PMC 53153. PMID 1722334. 
  8. ^ Braithwaite EK, Prasad R, Shock DD, Hou EW, Beard WA, Wilson SH (May 2005). "DNA polymerase lambda mediates a back-up base excision repair activity in extracts of mouse embryonic fibroblasts". J. Biol. Chem. 280 (18): 18469–75. doi:10.1074/jbc.M411864200. PMID 15749700. 
  9. ^ Beard WA, Prasad R, Wilson SH (2006). "Activities and mechanism of DNA polymerase beta". Meth. Enzymol. 408: 91–107. doi:10.1016/S0076-6879(06)08007-4. PMID 16793365. 
  10. ^ Kao HI, Henricksen LA, Liu Y, Bambara RA (April 2002). "Cleavage specificity of Saccharomyces cerevisiae flap endonuclease 1 suggests a double-flap structure as the cellular substrate". J. Biol. Chem. 277 (17): 14379–89. doi:10.1074/jbc.M110662200. PMID 11825897. 
  11. ^ Starcevic D, Dalal S, Sweasy JB (August 2004). "Is there a link between DNA polymerase beta and cancer?". Cell Cycle 3 (8): 998–1001. doi:10.4161/cc.3.8.1062. PMID 15280658. 
  12. ^ Farrington, S. M.; Tenesa, A; Barnetson, R; Wiltshire, A; Prendergast, J; Porteous, M; Campbell, H; Dunlop, M. G. (2005). "Germline susceptibility to colorectal cancer due to base-excision repair gene defects". The American Journal of Human Genetics 77 (1): 112–9. doi:10.1086/431213. PMC 1226182. PMID 15931596. 

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