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Nucleases are enzymes that hydrolytically cleave the phosphodiester backbone of DNA. Endonucleases cleave in the middle of chains, while exonucleases operate selectively by degrading from the end of the chain. Nucleases that act on both single- and double-stranded DNA are known. Restriction endonucleases are a special class that recognize and cleave specific sequences in DNA. Type II restriction endonucleases always…

Three types of natural methylation have been reported in DNA. Cytosine can be modified either on the ring to form 5-methylcytosine or on the exocyclic amino group to form N4-methylcytosine. Adenine may be modified to form N6-methyladenine. N4-methylcytosine and N6-methyladenine are found only in bacteria and archaea, whereas 5-methylcytosine is widely distributed. Special enzymes called DNA methyltransferases are responsible…

After a DNA molecule has been assembled, it may be chemically modified—sometimes deliberately by special enzymes called DNA methyltransferases and sometimes accidentally by oxidation, ionizing radiation, or the action of chemical carcinogens. DNA can also be cleaved and degraded by enzymes called nucleases.

DNA melting and reassociation can be monitored by measuring the absorption of ultraviolet (UV) light at a wavelength of 260 nanometres (billionths of a metre). When DNA is in a double-stranded conformation, absorption is fairly weak, but when DNA is single-stranded, the unstacking of the bases leads to an enhancement of absorption called hyperchromicity. Therefore, the…

Denaturation The strands of the DNA double helix are held together by hydrogen bonding interactions between the complementary base pairs. Heating DNA in solution easily breaks these hydrogen bonds, allowing the two strands to separate—a process called denaturation or melting. The two strands may reassociate when the solution cools, reforming the starting DNA duplex—a process called renaturation or hybridization. These…

Naturally occurring DNA molecules can be circular or linear. The genomes of single-celled bacteria and archaea (the prokaryotes), as well as the genomes of mitochondria and chloroplasts (certain functional structures within the cell), are circular molecules. In addition, some bacteria and archaea have smaller circular DNA molecules called plasmids that typically contain only a few genes. Many plasmids are readily transmitted from one cell to another.…

In 1953 James D. Watson and Francis H.C. Crick proposed a three-dimensional structure for DNA based on low-resolution X-ray crystallographic data and on Erwin Chargaff’s observation that, in naturally occurring DNA, the amount of T equals the amount of A and the amount of G equals the amount of C. Watson and Crick, who shared a Nobel Prize in 1962 for their efforts,…

DNA is a polymer of the four nucleotides A, C, G, and T, which are joined through a backbone of alternating phosphate and deoxyribose sugar residues. These nitrogen-containing bases occur in complementary pairs as determined by their ability to form hydrogen bonds between them. A always pairs with T through two hydrogen bonds, and G always pairs with C through three hydrogen bonds. The…

Nucleotides are synthesized from readily available precursors in the cell. The ribose phosphate portion of both purine and pyrimidine nucleotides is synthesized from glucose via the pentose phosphate pathway. The six-atom pyrimidine ring is synthesized first and subsequently attached to the ribose phosphate. The two rings in purines are synthesized while attached to the ribose phosphate during the assembly of adenine or guanine nucleosides. In…

Basic structure Nucleic acids are polynucleotides—that is, long chainlike molecules composed of a series of nearly identical building blocks called nucleotides. Each nucleotide consists of a nitrogen-containing aromatic base attached to a pentose (five-carbon) sugar, which is in turn attached to a phosphate group. Each nucleic acid contains four of five possible nitrogen-containing bases: adenine (A), guanine (G), cytosine (C), thymine (T), and uracil (U). A and G are categorized as purines, and…