Chimeric nucleases are an example of

engineered proteins Engineering is the practice of using natural science, mathematics, and the engineering design process to solve problems within technology, increase efficiency and productivity, and improve systems. Modern engineering comprises many subfiel ...

which must comprise a

DNA-binding domain A DNA-binding domain (DBD) is an independently folded protein domain that contains at least one structural motif that recognizes double- or single-stranded DNA. A DBD can recognize a specific DNA sequence (a recognition sequence) or have a gener ...

to give sequence specificity and a

nuclease In biochemistry, a nuclease (also archaically known as nucleodepolymerase or polynucleotidase) is an enzyme capable of cleaving the phosphodiester bonds that link nucleotides together to form nucleic acids. Nucleases variously affect single and ...

domain for

DNA Deoxyribonucleic acid (; DNA) is a polymer composed of two polynucleotide chains that coil around each other to form a double helix. The polymer carries genetic instructions for the development, functioning, growth and reproduction of al ...

cleavage.

DNA-binding domains

DNA-binding domains including the

basic helix-loop-helix Basic or BASIC may refer to: Science and technology * BASIC, a computer programming language * Basic (chemistry), having the properties of a base * Basic access authentication, in HTTP Entertainment * ''Basic'' (film), a 2003 film * Basic, one ...

zinc finger A zinc finger is a small protein structural motif that is characterized by the coordination of one or more zinc ions (Zn2+) which stabilizes the fold. The term ''zinc finger'' was originally coined to describe the finger-like appearance of a ...

helix-turn-helix Helix-turn-helix is a DNA-binding domain (DBD). The helix-turn-helix (HTH) is a major structural motif capable of binding DNA. Each monomer incorporates two alpha helix, α helices, joined by a short strand of amino acids, that bind to the majo ...

and

leucine zipper A leucine zipper (or leucine scissors) is a common three-dimensional structural motif in proteins. They were first described by Landschulz and collaborators in 1988 when they found that an enhancer binding protein had a very characteristic 30-amin ...

motifs have been used in construction of sequence-specific nucleases. Of these, zinc fingers have been suggested the most important due to their modularity, allowing construction of a tailor-made DNA-binding domain.

Nuclease domain

The nuclease domain is responsible for physical cleavage of DNA strands and may introduce either single stranded or double-stranded breaks.

FokI The restriction endonuclease Fok1, naturally found in ''Flavobacterium okeanokoites'', is a bacterial type IIS restriction endonuclease consisting of an N-terminal DNA-binding domain and a non sequence-specific DNA cleavage domain at the C-termi ...

is an example of a sequence-specific

endonuclease In molecular biology, endonucleases are enzymes that cleave the phosphodiester bond within a polynucleotide chain (namely DNA or RNA). Some, such as deoxyribonuclease I, cut DNA relatively nonspecifically (with regard to sequence), while man ...

whose non-specific nuclease domain introduces double stranded breaks and has been used in a variety of experiments including identification of high- and low-affinity binding sites of transcription factors ''in vitro'', to study recruitment of factors to promoter sites ''in vivo'' using protein position identification with a nuclease tail assay and to study proteins specific to interaction with DNA in the Z-DNA conformation (Durai et al., 2005 and references therein).

References

Engineered proteins Protein engineering Genetics experiments {{hydrolase-stub

DNA-binding domains

Nuclease domain

See also

References