Combinatorial targeting of ribbon-helix-helix artificial transcription factors to chimeric recognition sites

Massimiliano Zampini, Finbarr Hayes

Allbwn ymchwil: Cyfraniad at gyfnodolynErthygladolygiad gan gymheiriaid

4 Dyfyniadau(SciVal)
110 Wedi eu Llwytho i Lawr (Pure)


Artificial transcription factors (ATFs) are potent synthetic biology tools for modulating endogenous gene expression and precision genome editing. The ribbon-helix-helix (RHH) superfamily of transcription factors are widespread in bacteria and archaea. The principal DNA binding determinant in this family comprises a two-stranded antiparallel β-sheet (ribbons) in which a pair of eight-residue motifs insert into the major groove. Here, we demonstrate that ribbons of divergent RHH proteins are compact and portable elements that can be grafted into a common α-helical scaffold producing active ATFs. Hybrid proteins cooperatively recognize DNA sites possessing core tetramer boxes whose functional spacing is dictated by interactions between the α-helical backbones. These interactions also promote combinatorial binding of chimeras with different transplanted ribbons, but identical backbones, to synthetic sites bearing cognate boxes for each protein either in vitro or in vivo. The composite assembly of interacting hybrid proteins offers potential advantages associated with combinatorial approaches to DNA recognition compared with ATFs that involve binding of a single protein. Moreover, the new class of RHH ATFs may be utilized to re-engineer transcriptional circuits, or may be enhanced with affinity tags, fluorescent moieties or other elements for targeted genome marking and manipulation in bacteria and archaea.
Iaith wreiddiolSaesneg
Tudalennau (o-i)6673-6682
Nifer y tudalennau10
CyfnodolynNucleic Acids Research
Rhif cyhoeddi14
Dynodwyr Gwrthrych Digidol (DOIs)
StatwsCyhoeddwyd - Awst 2012

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