2VBN

Molecular basis of human XPC gene recognition and cleavage by engineered homing endonuclease heterodimers

Summary for 2VBN

• Entry DOI: 10.2210/pdb2vbn/pdb
• Related: 1AF5 1BP7 1G9Y 1G9Z 1MOW 1N3E 1N3F 1T9I 1T9J 1U0C 1U0D 2VBJ 2VBL 2VBO
• Descriptor: DNA ENDONUCLEASE I-CREI, 5'-D(*TP*TP*AP*GP*GP*AP*TP*CP*CP*TP *TP*CP*AP*AP)-3', 5'-D(*TP*CP*TP*GP*CP*CP*TP*TP*TP*TP *TP*TP*GP*AP)-3', ... (9 entities in total)
• Functional Keywords: uv-induced dna damage, cutting dna endonucleases, endonuclease, intron homing, ini3- 4_magnesium, plastid, nuclease, hydrolase, chloroplast, double strand break (dsb), homing endonucleases (hes)
• Biological source: CHLAMYDOMONAS REINHARDTII; More
• Cellular location: Plastid, chloroplast: P05725 2VBN
• Total number of polymer chains: 6
• Total formula weight: 49907.16

Authors

Redondo, P.,Prieto, J.,Munoz, I.G.,Alibes, A.,Stricher, F.,Serrano, L.,Arnould, S.,Perez, C.,Cabaniols, J.P.,Duchateau, P.,Paques, F.,Blanco, F.J.,Montoya, G. (deposition date: 2007-09-14, release date: 2008-10-28, Last modification date: 2023-12-13)

Primary citation

Redondo, P.,Prieto, J.,Munoz, I.G.,Alibes, A.,Stricher, F.,Serrano, L.,Cabaniols, J.P.,Daboussi, F.,Arnould, S.,Perez, C.,Duchateau, P.,Paques, F.,Blanco, F.J.,Montoya, G.
Molecular Basis of Xeroderma Pigmentosum Group C DNA Recognition by Engineered Meganucleases
Nature, 456:107-, 2008

PubMed Abstract: Xeroderma pigmentosum is a monogenic disease characterized by hypersensitivity to ultraviolet light. The cells of xeroderma pigmentosum patients are defective in nucleotide excision repair, limiting their capacity to eliminate ultraviolet-induced DNA damage, and resulting in a strong predisposition to develop skin cancers. The use of rare cutting DNA endonucleases-such as homing endonucleases, also known as meganucleases-constitutes one possible strategy for repairing DNA lesions. Homing endonucleases have emerged as highly specific molecular scalpels that recognize and cleave DNA sites, promoting efficient homologous gene targeting through double-strand-break-induced homologous recombination. Here we describe two engineered heterodimeric derivatives of the homing endonuclease I-CreI, produced by a semi-rational approach. These two molecules-Amel3-Amel4 and Ini3-Ini4-cleave DNA from the human XPC gene (xeroderma pigmentosum group C), in vitro and in vivo. Crystal structures of the I-CreI variants complexed with intact and cleaved XPC target DNA suggest that the mechanism of DNA recognition and cleavage by the engineered homing endonucleases is similar to that of the wild-type I-CreI. Furthermore, these derivatives induced high levels of specific gene targeting in mammalian cells while displaying no obvious genotoxicity. Thus, homing endonucleases can be designed to recognize and cleave the DNA sequences of specific genes, opening up new possibilities for genome engineering and gene therapy in xeroderma pigmentosum patients whose illness can be treated ex vivo.

PubMed: 18987743
DOI: 10.1038/NATURE07343

Experimental method

X-RAY DIFFRACTION (1.9 Å)