TY - JOUR
T1 - TAT-mediated delivery of a DNA repair enzyme to skin cells rapidly initiates repair of UV-induced DNA damage
AU - Johnson, Jodi L.
AU - Lowell, Brian C.
AU - Ryabinina, Olga P.
AU - Lloyd, R. Stephen
AU - McCullough, Amanda K.
N1 - Funding Information:
This research was funded in part by NIH R01 ES04091 and Restoration Genetics, STTR Grant R41 CA114923. Dr Jodi L. Johnson's salary was supported by the Ruth L. Kirschstein National Research Service Training Grant Award ES007060-28 from Oregon State University's Department of Environmental and Molecular Toxicology, and some research supplies were provided by a Dermatology Foundation Research Grant. We acknowledge Dr Yih-Tai Chen at OTRADI for his expertise with automated microscopy. Dr Mihail Iordanov provided the E6/E7 immortalized skin fibroblasts. We thank Lauriel Earley, James Lagowski, and Drs Anuradha Kumari and Irina G. Minko for insightful discussions concerning these investigations and critically reading the manuscript.
PY - 2011/3
Y1 - 2011/3
N2 - UV light causes DNA damage in skin cells, leading to more than one million cases of non-melanoma skin cancer diagnosed annually in the United States. Although human cells possess a mechanism (nucleotide excision repair) to repair UV-induced DNA damage, mutagenesis still occurs when DNA is replicated before repair of these photoproducts. Although human cells have all the enzymes necessary to complete an alternate repair pathway, base excision repair (BER), they lack a DNA glycosylase that can initiate BER of dipyrimidine photoproducts. Certain prokaryotes and viruses produce pyrimidine dimer-specific DNA glycosylases (pdgs) that initiate BER of cyclobutane pyrimidine dimers (CPDs), the predominant UV-induced lesions. Such a pdg was identified in the Chlorella virus PBCV-1 and termed Cv-pdg. The Cv-pdg protein was engineered to contain a nuclear localization sequence (NLS) and a membrane permeabilization peptide (transcriptional transactivator, TAT). Here, we demonstrate that the Cv-pdg-NLS-TAT protein was delivered to repair-proficient keratinocytes and fibroblasts, and to a human skin model, where it rapidly initiated removal of CPDs. These data suggest a potential strategy for prevention of human skin cancer.
AB - UV light causes DNA damage in skin cells, leading to more than one million cases of non-melanoma skin cancer diagnosed annually in the United States. Although human cells possess a mechanism (nucleotide excision repair) to repair UV-induced DNA damage, mutagenesis still occurs when DNA is replicated before repair of these photoproducts. Although human cells have all the enzymes necessary to complete an alternate repair pathway, base excision repair (BER), they lack a DNA glycosylase that can initiate BER of dipyrimidine photoproducts. Certain prokaryotes and viruses produce pyrimidine dimer-specific DNA glycosylases (pdgs) that initiate BER of cyclobutane pyrimidine dimers (CPDs), the predominant UV-induced lesions. Such a pdg was identified in the Chlorella virus PBCV-1 and termed Cv-pdg. The Cv-pdg protein was engineered to contain a nuclear localization sequence (NLS) and a membrane permeabilization peptide (transcriptional transactivator, TAT). Here, we demonstrate that the Cv-pdg-NLS-TAT protein was delivered to repair-proficient keratinocytes and fibroblasts, and to a human skin model, where it rapidly initiated removal of CPDs. These data suggest a potential strategy for prevention of human skin cancer.
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U2 - 10.1038/jid.2010.300
DO - 10.1038/jid.2010.300
M3 - Article
C2 - 20927123
AN - SCOPUS:79951484657
SN - 0022-202X
VL - 131
SP - 753
EP - 761
JO - Journal of Investigative Dermatology
JF - Journal of Investigative Dermatology
IS - 3
ER -