Sains Malaysiana 29: 19-31 (2000)                                                                                                                Sains Hayat /

Life Sciences

DNA Repair at the Gene Level in

Trichothiodystrophy Cells

Sheila Nathan

Centre for Gene Analysis and Technology

Faculty of Science & Technology

Universiti Kebangsaan Malaysia

43600 Bangi Selangor D. E, Malaysia

Lynne Mayne

Trafford Centre for Graduate Medical Research

University of Sussex, Brighton BN1 9RY

England 31

ABSTRACT

Cells from trichothiodystrophy (TTD) group 3 patients are non-hypersensitive to uv irradiation. These cells perform normal genome repair of pyrimidine dimers but fail to excise 6-4 photoproducts and concomitantly, are unable to restore RNA synthesis levels following uv irradiation. This pointed to a defect in gene-specific repair and this study was undertaken to examine repair of dimers at the gene-level. The results indicated a defect in gene-specific repair of dimers at early times post-irradiation due to the inability of the cells to repair dimers from the transcribed strand of the active gene. These findings indicate that TTD group 3 cells are unable to remove DNA lesions preferen­tially and this might contribute to other inherent symptoms of these patients.

ABSTRAK

Sel daripada pesakit trikotiodistrofi (TTD) kumpulan 3 tidak sensitif terhadap sinaran ultraungu. Sel ini mampu membaikpulih dimer pirimidin keseluruhan genom secara normal tetapi gagal memperbaiki fotoproduk 6-4 dan ini menyebabkan terencatnya pemulihan sintesis RNA selepas sinaran UV. Keadaan ini menunjukkan kecacatan dalam pembaikpulih spesifik-gen. Oleh itu, projek ini bertujuan untuk mengkaji pembaikan dimer pada peringkat gen. Keputusan yang diperolehi menunjukkan kecacatan pembaikpulihan pada peringkat awal selepas sinaran disebabkan oleh ketidakupayaan sel untuk mengeluarkan dimer dari rantai tertranskripsi gen aktif. Ini menandakan sel TTD kumpulan 3 tidak berupaya mengeluarkan lesi DNA secara terpilih dan ini mungkin menyumbang kepada simptom lain pada pesakit TTD.

RUJUKAN/REFERENCES

Berkvens Th. M., Gerritsen E.J.A., Oldenburg M., Van der Eb A.J. & Meera Khan p. 1987.  Severe combined immunodeficiency due to a homozygous 3.2 kb deletion spanning the promoter and first exon of the adenosine deaminase gene. Nucl. Acid. Res. 15: 9365-9378.

Biernat, J., Gobel U.B. & Kostel H. 1989.  New bacteriophage vectors for the large-scale production of single-stranded DNA. J. Biochem. And Biophys. Methods. 19: 155-168.

Bohr V.A., C.A. Smith D.S. Okumoto & P.C. Hanawalt (1985).  DNA repair in a active gene: Removal of pyrimidine dimmers from the DHFR gene of CHO cells is much more efficient than in the genome overall. Cell 40: 359-369.

Bohr V.A. 1991.  Gene specific damage and repair after treatment of cells with UV and chemotherapeutical agents. Adv Exp Med Biol. 283: 225-233.

Broughton B.C., Lehmann A.R., Harcourt S.A., Arlett C.F., Sarasin A., Kleijer W.J., Beemer F.A., Naim R. & Mitchell, D.L. 1990.  Relationships between pyrimidine dimmers, 6-4 photoproducts, repair synthesis and cell survival: Studies using cells from patients with trichothiodystrophy. Mutation Res. 235: 33-40.

Coin F., Marinoni J.C. & Egly J.M. 1998. Mutations in XPD helicase prevent its interaction and regulation by p44, another subunit of TFIIH, resulting in Xero­derma pigmentosum and trichothiodystrophy phenotypes. Pathol. Bio. 46: 679-­680.

Feinberg A.P., & Vogel stein B. 1983. A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem. 132: 6.

Guo Z., Heydari A., & Richardson A. (1998). Nucleotide excision repair of actively transcribed versus nontranscribed DNA in rat hepatocytes: effect of age and dietary restriction. Exp Cell Res 245: 228-38

Hofker M.H., Van Ommen G.J.B., Bakker E., Burmeister M. & Pearson P.L. 1986. Development of additional RFLP probes near the locus for Duchenne muscular dystrophy by cosmid cloning of the DSX84 (754) locus. Hum. Genet. 74: 270­-274.

Kantor G.J., Barsalou L.S. & Hanawalt P.C. 1990. Selective repair of specific chromatin domains in UV-irradiated cells from xeroderma pigmentosum complementation group C. Mutation Res. 235: 171-180.

Lattier D.L., States J.C., Hutton J.J. & Wiginton D.A. 1989. Cell type-specific transcriptional regulation of the human adenosine deaminase gene. Nucl. Acid Res. 17: 1061-1076.

Lehmann A.R, Arlett C.F., Broughton B.C., Harcourt S.A., Steingrimsdottir H., Stefanini M., Taylor M.R, Natarajan A.T., Green S., King M.D., Mackie R.M.M., Stephenson J.B.P. & Tolmie J.L. 1988. TTD, a human DNA repair disorder with heterogeneity in the response to UV light. Cancer Res. 48: 6090­-6096.

Nakabeppu Y., Yamashiata K. & Sekiguchi M. 1982. Purification and characteriza­tion of normal and mutant forms of T4 endonuclease V. J. Bio. Chem. 257: 2556-2662.

Price V.H., adorn RB., Ward W.H. & Jones F.T. 1980. Trichothiodystrophy: Sulphur-deficient brittle hair as a marker for a neuroectodermal symptom complex. Arch. Dermatol. 116: 1375-1384.

Tijsterman M., De Pril R., Tasseron-De Jong J.G. & Brouwer J. 1999. RNA polymerase II transcription suppresses nucleosomal modulation of UV induced (6-4) photoproduct and cyclobutane pyrimidine dimer repair in yeast. Mol. Cell. Bio. 19: 934-940.

Van Hoffen A., Venema J., Meschini R., Van Zeeland A.A. & Mullenders L.H. 1995. Transcription-coupled repair removes both cyclobutane pyrimidine dimers and 6­4 photoproducts with equal efficiency and in a sequential way from transcribed DNA in xeroderma pigmentosum group C fibroblasts. EMBOJ. 14: 360-7.

Venema J., Mullenders L.H., Natarajan A.T., Van Zeeland A.A. & Mayne L.V. 1990a. The genetic defect in Cockayne syndrome is associated with a defect in repair of UV-induced DNA damage in transcriptionally active DNA. Proc. Natl. Acad. Sci. USA. 87: 4707-11.

Venema J., Van Hoffen A., Natarajan A.T., Van Zeeland A.A. & Mullenders L.H. 1990b. The residual repair capacity of xeroderma pigmentosum complementation group C fibroblasts is highly specific for transcriptionally active DNA. Nucl. Acids Res. 18: 443-448.

Weeda G., Eveno E., Donker I., Vermeulen W., Chevallier-Lagente 0., Taieb A., Stary A., Hoeijmakers J.H., Mezzina M. & Sarasin A. 1997. A mutation in the XPBlERCC3 DNA repair transcription gene, associated with trichothiodystrophy. Am. J. Hum. Genet. 60: 320-329.

WiIl C.L. & Dolnick B.J. 1986. Fluorouracil augmentation of dihydrofolate reductase agene transcripts containing intervening sequences in methotrexate-resistance KB cells. Mol. Pharmacol. 29: 643-648.

Yang I.K., Masters J.N. & Atardi G. 1984. Human dihydrofolate reductase gene organization. J. Mol. BioI. 176: 169-187.