Mutagenesis may occur endogenously, for example through spontaneous hydrolysis, or through normal cellular processes that can generate reactive oxygen species and DNA adducts, or through error in replication and repair.[15] Mutagenesis may also arise as a result of the presence of environmental mutagens that induces changes to the DNA. The mechanism by which mutation arises varies according to the causative agent, the mutagen, involved. Most mutagens act either directly, or indirectly via mutagenic metabolites, on the DNA producing lesions. Some however may affect the replication or chromosomal partition mechanism, and other cellular processes.
Many chemical mutagens require biological activation to become mutagenic. An important group of enzymes involved in the generation of mutagenic metabolites is cytochrome P450.[16] Other enzymes that may also produce mutagenic metabolites include glutathione S-transferase and microsomal epoxide hydrolase. Mutagens that are not mutagenic by themselves but require biological activation are called promutagens.
Many mutations arise as a result of problems caused by the DNA lesions during replication, resulting in errors in replication. In bacteria, extensive damage to the DNA due to mutagens results in single-stranded DNA gaps during replication. This induces the SOS response, an emergency repair process that is also error-prone, thereby generating mutations. In mammalian cells, stalling of replication at a damaged sites induces a number of rescue mechanisms that help bypass DNA lesions, but which also may result in errors. The Y family of DNA polymerases specialize in DNA lesion bypass in a process termed translesion synthesis (TLS) whereby these lesion-bypass polymerases replace the stalled high-fidelity replicative DNA polymrase, transits the lesion and extend the DNA until the lesion has been passed so that normal replication can resume. These processes may be error-prone or error-free.
Spontaneous hydrolysis
DNA is not entirely stable in aqueous solution. Under physiological conditions the glycosidic bond may be hydrolyzed spontaneously and 10,000 purine sites in DNA are estimated to be depurinated each day in a cell.[15] Numerous DNA repair pathway exist for the DNA, however, if the apurinic site failed to be repaired, misincorporation of nucleotide may occur during replication. Adenine is preferentially incorporated by DNA polymerases in an apurinic site.
Cytidine may also become deaminated to uridine at one five-hundredth of the rate of depurination and can result in G to A transition. Eukaryotic cells also contains 5-methylcytosine, thought to be involved in the control of gene transcription, which can become deaminated into thymine.
Modification of bases
Bases may be modified endogenously by normal cellular molecules. For example DNA may be methylated by S-adenosylmethionine, and glycosylated by reducing sugars.
Many compounds, such as PAHs, aromatic amines, aflatoxin and pyrrolizidine alkaloids, may form reactive oxygen species catalyzed by cytochrome P450. These metabolites form adducts with the DNA, which can cause errors in replication, and the bulky aromatic adducts may form stable intercalation between bases and block replication. The adducts may also induce conformational changes in the DNA. Some adducts may also result in the depurination of the DNA,[17] it is however uncertain how significant such depurination as caused by the adducts is in generating mutation.[18]
Some alkylating agents such as N-Nitrosamines may also require the catalytic reaction of cytochrome-P450 for the formation of a reactive alkyl cation. Alkylation and arylation of bases can cause errors in replication. N7 and O6 of guanine and the N3 and N7 of adenine are most susceptible to attack; while N7-guanine adducts, which form the bulk of DNA adducts, appear to be non-mutagenic, alkylation at O6 of guanine is harmful because excision repair of O6-adduct of guanine may be poor in some tissues such as the brain.[19] The O6 methylation of guanine can result in G to A transition, while O4-methylthymine can be mispaired with guanine. The type of the mutation generated however may be dependent on the size and type of the adduct as well as the DNA sequence.[20]
Ionizing radiations and reactive oxygen species often oxidize guanine to produce 8-oxoguanine.
Crosslinking
Main article: Crosslinking of DNA
Some alkylating agents may produce crosslinking of DNA. Some natural occurring chemicals may also promotes crosslinking, such as psoralens after activation by UV radiation, and nitrous acid. Interstrand cross-linking is more damaging as it blocks replication and transcription and can cause chromosomal breakages and rearrangements. Some crosslinkers such as cyclophosphamide, mitomycin C and cisplatin are used as anticancer chemotherapeutic because their high degree of toxicity to proliferating cells.
Dimerization
UV radiation promotes the formation of a cyclobutyl ring between adjacent thymines, resulting in the formation of pyrimidine dimers.[21] In human skin cells, thousands of dimers may be formed in a day due to normal exposure to sunlight. DNA polymerase η may help bypass these lesions in an error-free manner;[22] however, individuals with defective DNA repair function, such as sufferers of Xeroderma pigmentosum, are sensitive to sunlight and may be prone to skin cancer.
Ethidium intercalated between two adenine-thymine base pairs.
Intercalation between bases
The planar structure of chemicals such as ethidium bromide and proflavine allows them to insert between bases in DNA, and cause frameshift mutation. The intercalation into DNA of anthracyclines such as daunorubicin and doxorubicin interferes with the functioning of the enzyme topoisomerase II, blocking replication as well as causing mitotic homologous recombination.
Arrows indicates chromosomal breakages due to DNA damage
Backbone damage
Ionizing radiations may produce highly reactive free radicals that can break the bonds in the DNA. Double-stranded breakages are especially damaging and hard to repair, producing translocation and deletion of part of a chromosomes. Alkylating agents like mustard gas may also cause breakages in the DNA backbone. Oxidative stress may also generate highly reactive oxygen species that can damage the DNA. Incorrect repair of other damages induced by the highly reactive species can also lead to mutations.
Insertional mutagenesis
Main article: Insertional mutagenesis
Transposon and virus may insert DNA sequence into coding region or functional elements of a gene and result in inactivation of the gene.
Error in replication
While most mutagens produce effects that ultimately result in error in replication, some mutagens may affect directly the replication process. Base analog such as 5-bromouracil may substitute for thymine in replication. Some metals such as cadmium, chromium, and nickel may alter the fidelity of DNA replication.