TAT-mediated delivery of a DNA repair enzyme to skin cells rapidly initiates repair of UV-induced DNA damage

Jodi L. Johnson; Brian C. Lowell; Olga P. Ryabinina; R. Stephen Lloyd; Amanda K. McCullough

doi:10.1038/jid.2010.300

TAT-mediated delivery of a DNA repair enzyme to skin cells rapidly initiates repair of UV-induced DNA damage

Jodi L. Johnson, Brian C. Lowell, Olga P. Ryabinina, R. Stephen Lloyd, Amanda K. McCullough

Pathology

Research output: Contribution to journal › Article › peer-review

21 Scopus citations

Abstract

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.

Original language	English (US)
Pages (from-to)	753-761
Number of pages	9
Journal	Journal of Investigative Dermatology
Volume	131
Issue number	3
DOIs	https://doi.org/10.1038/jid.2010.300
State	Published - Mar 2011

Funding

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.

ASJC Scopus subject areas

Biochemistry
Molecular Biology
Dermatology
Cell Biology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1038/jid.2010.300

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title = "TAT-mediated delivery of a DNA repair enzyme to skin cells rapidly initiates repair of UV-induced DNA damage",

abstract = "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.",

author = "Johnson, {Jodi L.} and Lowell, {Brian C.} and Ryabinina, {Olga P.} and Lloyd, {R. Stephen} and McCullough, {Amanda K.}",

note = "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. ",

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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

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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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TAT-mediated delivery of a DNA repair enzyme to skin cells rapidly initiates repair of UV-induced DNA damage

Abstract

Funding

ASJC Scopus subject areas

UN SDGs

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