Mutagenesis in mammalian cells induced by triple helix formation and transcription-coupled repair

Science

Volumn 271, Issue 5250, 1996, Pages 802-805

  Wang, Gan ^a   Seidman, Michael M ^b   Glazer, Peter M ^a  

^a YALE UNIVERSITY SCHOOL OF MEDICINE   (United States)

^b Oncor Inc   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

OLIGONUCLEOTIDE;

ANIMAL CELL; ARTICLE; COCKAYNE SYNDROME; CONTROLLED STUDY; DNA REPAIR; GENETIC STABILITY; GENETIC TRANSCRIPTION; HUMAN; HUMAN CELL; MAMMAL CELL; MUTAGENESIS; NONHUMAN; PRIORITY JOURNAL; SIMIAN VIRUS 40; TRIPLE HELIX; XERODERMA PIGMENTOSUM;

ANIMALIA; MAMMALIA; SIMIAE; SIMIAN VIRUS; SIMIAN VIRUS 40;

EID: 0030059301     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.271.5250.802     Document Type: Article

References (53)

1. P A. Beal and P. B. Dervan, Science 251, 1360 (1991), A. G. Letai, M A Palladino, E. Fromm, V RIZZO, J. R Fresco, Biochemistry 27, 9108 (1988), C. Helene, Curr. Opin. Biotechnol 4, 29 (1993); H E. Moser and P. B. Dervan, Science 238, 645 (1987), D. Praseuth et al., Proc. Natl Acad Sci. U.S A 85, 1349 (1988); M Cooney, G Czernuszewicz, E H Postel, S. J. Flint, M. E. Hogan, Science 241, 456 (1988)
2. P A. Beal and P. B. Dervan, Science 251, 1360 (1991), A. G. Letai, M A Palladino, E. Fromm, V RIZZO, J. R Fresco, Biochemistry 27, 9108 (1988), C. Helene, Curr. Opin. Biotechnol 4, 29 (1993); H E. Moser and P. B. Dervan, Science 238, 645 (1987), D. Praseuth et al., Proc. Natl Acad Sci. U.S A 85, 1349 (1988); M Cooney, G Czernuszewicz, E H Postel, S. J. Flint, M. E. Hogan, Science 241, 456 (1988)
3. P A. Beal and P. B. Dervan, Science 251, 1360 (1991), A. G. Letai, M A Palladino, E. Fromm, V RIZZO, J. R Fresco, Biochemistry 27, 9108 (1988), C. Helene, Curr. Opin. Biotechnol 4, 29 (1993); H E. Moser and P. B. Dervan, Science 238, 645 (1987), D. Praseuth et al., Proc. Natl Acad Sci. U.S A 85, 1349 (1988); M Cooney, G Czernuszewicz, E H Postel, S. J. Flint, M. E. Hogan, Science 241, 456 (1988)
4. P A. Beal and P. B. Dervan, Science 251, 1360 (1991), A. G. Letai, M A Palladino, E. Fromm, V RIZZO, J. R Fresco, Biochemistry 27, 9108 (1988), C. Helene, Curr. Opin. Biotechnol 4, 29 (1993); H E. Moser and P. B. Dervan, Science 238, 645 (1987), D. Praseuth et al., Proc. Natl Acad Sci. U.S A 85, 1349 (1988); M Cooney, G Czernuszewicz, E H Postel, S. J. Flint, M. E. Hogan, Science 241, 456 (1988)
5. P A. Beal and P. B. Dervan, Science 251, 1360 (1991), A. G. Letai, M A Palladino, E. Fromm, V RIZZO, J. R Fresco, Biochemistry 27, 9108 (1988), C. Helene, Curr. Opin. Biotechnol 4, 29 (1993); H E. Moser and P. B. Dervan, Science 238, 645 (1987), D. Praseuth et al., Proc. Natl Acad Sci. U.S A 85, 1349 (1988); M Cooney, G Czernuszewicz, E H Postel, S. J. Flint, M. E. Hogan, Science 241, 456 (1988)
6. P A. Beal and P. B. Dervan, Science 251, 1360 (1991), A. G. Letai, M A Palladino, E. Fromm, V RIZZO, J. R Fresco, Biochemistry 27, 9108 (1988), C. Helene, Curr. Opin. Biotechnol 4, 29 (1993); H E. Moser and P. B. Dervan, Science 238, 645 (1987), D. Praseuth et al., Proc. Natl Acad Sci. U.S A 85, 1349 (1988); M Cooney, G Czernuszewicz, E H Postel, S. J. Flint, M. E. Hogan, Science 241, 456 (1988)
7. G Duval-Valentin, N. T. Thuong, C. Helene, Proc Natl. Acad. Sci. U.S.A. 89, 504 (1992); N. H Ing et al., Nucleic Acids Res. 21, 2789 (1993); L J Maher III, B. Wold, P B Dervan, Science 245, 725 (1989), E H Postel, S. J. Flint, D. J. Kessler, M. E. Hogan, Proc Natl. Acad. Sci. USA 88, 8227 (1991), S. L. Young, S. H. Krawczyk, M D. Matteucci, J. J Toole, ibid., p 10023; M. Grigoriev et al , ibid. 90, 3501 (1993).
8. G Duval-Valentin, N. T. Thuong, C. Helene, Proc Natl. Acad. Sci. U.S.A. 89, 504 (1992); N. H Ing et al., Nucleic Acids Res. 21, 2789 (1993); L J Maher III, B. Wold, P B Dervan, Science 245, 725 (1989), E H Postel, S. J. Flint, D. J. Kessler, M. E. Hogan, Proc Natl. Acad. Sci. USA 88, 8227 (1991), S. L. Young, S. H. Krawczyk, M D. Matteucci, J. J Toole, ibid., p 10023; M. Grigoriev et al , ibid. 90, 3501 (1993).
9. G Duval-Valentin, N. T. Thuong, C. Helene, Proc Natl. Acad. Sci. U.S.A. 89, 504 (1992); N. H Ing et al., Nucleic Acids Res. 21, 2789 (1993); L J Maher III, B. Wold, P B Dervan, Science 245, 725 (1989), E H Postel, S. J. Flint, D. J. Kessler, M. E. Hogan, Proc Natl. Acad. Sci. USA 88, 8227 (1991), S. L. Young, S. H. Krawczyk, M D. Matteucci, J. J Toole, ibid., p 10023; M. Grigoriev et al , ibid. 90, 3501 (1993).
10. G Duval-Valentin, N. T. Thuong, C. Helene, Proc Natl. Acad. Sci. U.S.A. 89, 504 (1992); N. H Ing et al., Nucleic Acids Res. 21, 2789 (1993); L J Maher III, B. Wold, P B Dervan, Science 245, 725 (1989), E H Postel, S. J. Flint, D. J. Kessler, M. E. Hogan, Proc Natl. Acad. Sci. USA 88, 8227 (1991), S. L. Young, S. H. Krawczyk, M D. Matteucci, J. J Toole, ibid., p 10023; M. Grigoriev et al , ibid. 90, 3501 (1993).
11. G Duval-Valentin, N. T. Thuong, C. Helene, Proc Natl. Acad. Sci. U.S.A. 89, 504 (1992); N. H Ing et al., Nucleic Acids Res. 21, 2789 (1993); L J Maher III, B. Wold, P B Dervan, Science 245, 725 (1989), E H Postel, S. J. Flint, D. J. Kessler, M. E. Hogan, Proc Natl. Acad. Sci. USA 88, 8227 (1991), S. L. Young, S. H. Krawczyk, M D. Matteucci, J. J Toole, ibid., p 10023; M. Grigoriev et al , ibid. 90, 3501 (1993).
12. G Duval-Valentin, N. T. Thuong, C. Helene, Proc Natl. Acad. Sci. U.S.A. 89, 504 (1992); N. H Ing et al., Nucleic Acids Res. 21, 2789 (1993); L J Maher III, B. Wold, P B Dervan, Science 245, 725 (1989), E H Postel, S. J. Flint, D. J. Kessler, M. E. Hogan, Proc Natl. Acad. Sci. USA 88, 8227 (1991), S. L. Young, S. H. Krawczyk, M D. Matteucci, J. J Toole, ibid., p 10023; M. Grigoriev et al , ibid. 90, 3501 (1993).
13. S. A Strobel, L A. Doucette-Stamm, L. Riba, D E. Housman, P. B. Dervan, Science 254, 1639 (1991).
14. P A. Havre, E. J Gunther, F. P Gasparro, P. M. Glazer, Proc Natl. Acad. Sci. U.S.A 90, 7879 (1993); P. A. Havre and P. M. Glazer, J Virol 67, 7324 (1993).
15. P A. Havre, E. J Gunther, F. P Gasparro, P. M. Glazer, Proc Natl. Acad. Sci. U.S.A 90, 7879 (1993); P. A. Havre and P. M. Glazer, J Virol 67, 7324 (1993).
16. G. Wang, D D. Levy, M. M. Seidman, P M. Glazer, Mol. Cell Biol. 15, 1759 (1995).
17. Oligonuoleotide sequences were AG10, 5′-AGGAAG-GGGG-3′, AG20, 5′-AGGAAGGGGGGGGTGGTG-GG-3′; AG30, 5′-AGGAAGGGGGGGGTGGTGGGG-GAGGGGGAG-3′, and Mix30, 5′-AGTCAGTCAGTCA-GTCAGTCAGTCAGTCAG-3′. Oligonucleotides were synthesized by J. Flory (W. M. Keck Biotechnology Resource Center, Yale University) with materials from Glen Research (Sterling, VA) The polypurine triplex target site was base pairs 167 to 196 of supFG1 (see Fig. 1) (5).
18. 2) were transfected by adding to the culture medium (10 ml volume) pSupFG1 DNA (5 μg) premixed with cationic liposomes (50 μg) (DOTAP, Boehringer Mannheim, Indianapolis, IN) After 12 hours, the cell monolayers were washed three times and fresh medium containing the selected oligonucleotide at a concentration of 2 μM was added
19. The oligonucleotides were not conjugated to any mutagen but were modified to resist nuclease-mediated degradation by incorporation of either a 3′ propylamine group or phosphorothioate internucleoside linkages. Similar results were obtained with either modification.
20. Isolation of the SV40 vector DNA and transformation of bacteria for genetic analysis of the supFG1 gene were performed as described (5).
21. Sequencing of the supFG1 gene mutations was performed directly from the plasmid vector DNA with a primer complementary to a region in the β-lactamase gene adjacent to the supFG1 gene, as described (5).
22. XPA cells from patient XP12BE were obtained from the NIGMS Human Genetic Mutant Cell Repository (Camden, NJ, repository number GM04429E). These cells were SV40-transformed fibroblasts from a patient with XPA. CSB cells (CS2BE, repository number GM01098B) were SV40-transformed flbroblasts from a patient with CSB Normal fibroblasts (repository number GM00637F) were SV40-transformed cells from an apparently normal donor. Transformed XPA fibroblasts from patient XP2OS and the XP2OS cells transfected with a vector expressing XPA cDNA (XP2OS-pCAH19WS) were obtained from K. Kraemer (NIH) (13). The cells were grown in DMEM supplemented with 10% FCS (Life Technologies).
23. C. H. Park and A Sancar, Proc. Natl. Acad. Sci. U S A 91, 5017 (1994); C. J. Jones and R D Wood, Biochemistry 32, 12096 (1993).
24. C. H. Park and A Sancar, Proc. Natl. Acad. Sci. U S A 91, 5017 (1994); C. J. Jones and R D Wood, Biochemistry 32, 12096 (1993).
25. D. D. Levy, M. Saijo, K. Tanaka, K H. Kraemer, Carcinogenesis 16, 1557 (1995).
26. C Troelstra et al., Cell 71, 939 (1992)
27. 2 flasks, were transfected with 20 μg of pSupFG1 and 10 μg of pSLME6(+) DNA (14) premixed with cationic liposomes (100 μg) (DOTAP; Boehnnger Mannheim). After 12 hours, the monolayer was washed with phosphate-buffered saline and fresh medium containing the selected oligonucleotides at 2 μM was added. Forty-eight hours later, the vector DNA was isolated for analysis as described (5, 10) Extracted DNA was subjected to digestion with Sal I to eliminate any persisting pSLME6 plasmid DNA and with Dpn I to eliminate any unreplicated pSupFG1 vector molecules. Plasmid DNA was analyzed from all of the white colonies to confirm that the colonies arose from pSupFG1 mutants and not from pSLME6(I)
28. Transcription of supF gene sequences in COS cells was examined by Northern blot analysis of total cellular RNA by the method of Chirgwin et al. [J H. Chirgwin, D E. Przybyla, R. J MacDonald, W. J Rutter, Biochemistry 18, 5294 (1979)]. RNA was resolved by electrophoresis through formaldehyde agarose gels and transferred to nylon filters Synthetic oligonucleotides of 30 bp, matching the sequences of the sense and antisense strands at the 5′ end of the supF gene, were used as probes Hybridization was done at 42°C in 20% formamide, 5x standard saline citrate (SSC), 1x Denhardt's solution, 0 05 M sodium phosphate (pH 7 2), 0.1% SDS, and 1 mg/ml of salmon sperm DNA, followed by autoradiography
29. R. D Wood, P. Robins, T Lindahl, Cell 53, 97 (1988).
30. Sibghat-Ullah, I Husain, W Carlton, A Sancar, Nucleic Acids Res. 17, 447 (1989), M. K K. Shivji, M K Kenny, R D Wood, Cell 69, 367 (1992); J. T. Reardon et al., Nucleic Acids Res 19, 4623 (1991)
31. Sibghat-Ullah, I Husain, W Carlton, A Sancar, Nucleic Acids Res. 17, 447 (1989), M. K K. Shivji, M K Kenny, R D Wood, Cell 69, 367 (1992); J. T. Reardon et al., Nucleic Acids Res 19, 4623 (1991)
32. Sibghat-Ullah, I Husain, W Carlton, A Sancar, Nucleic Acids Res. 17, 447 (1989), M. K K. Shivji, M K Kenny, R D Wood, Cell 69, 367 (1992); J. T. Reardon et al., Nucleic Acids Res 19, 4623 (1991)
33. 32P]dCTP, additionally supplemented as described (77), and incubated for 3 hours at 30°C The DNA was extracted with phenol-chloroform, concentrated by filtration with a Centricon 100 filter (Amicon, Beverly, MA), and linearized with Eco RI The samples were analyzed by 0.7% agarose gel electrophoresis, ethidium bromide staining, and autoradiography.
34. 32P]guanosine 5′-triphosphate at 30°C for 1 hour. RNA products were visualized by electrophoresis on a 4% denaturing polyacrylamide gel containing 7 M urea, followed by autoradiography.
35. P. C. Hanawalt, Science 266, 1957 (1994).
36. A. Datta and S Jinks-Robertson, ibid. 268, 1616 (1995)
37. S K. Quah, R. C von Borstel, P. J. Hastings, Genetics 96, 819 (1980), C. W. Lawrence, T. O'Brien, J Bond, Mol Gen. Genet 195, 487 (1984) L Prakash, Genetics 83, 285 (1976).
38. S K. Quah, R. C von Borstel, P. J. Hastings, Genetics 96, 819 (1980), C. W. Lawrence, T. O'Brien, J Bond, Mol Gen. Genet 195, 487 (1984) L Prakash, Genetics 83, 285 (1976).
39. S K. Quah, R. C von Borstel, P. J. Hastings, Genetics 96, 819 (1980), C. W. Lawrence, T. O'Brien, J Bond, Mol Gen. Genet 195, 487 (1984) L Prakash, Genetics 83, 285 (1976).
40. J S. Lee, G. D. Burkhoider, L. J. P Latimer, B. L. Haug, R. P. Braun, Nucleic Acids Res. 15, 1047 (1987); S. M Mirkin and M D Frank-Kamenetski, Annu. Rev Biomol Struct 23, 541 (1994), M. J. Ulrich, W. J. Gray, T. J. Ley, J. Biol Chem. 267, 18649 (1992), Y. Kohwi and T. Kohwi-Shigematsu, Proc. Natl. Acad Sci. U.S.A. 85, 3781 (1988); A J. Kinniburgh, Nucleic Acids Res 17, 7771 (1989); D Sen and W. Gilbert, Nature 334, 364 (1988), W I Sundquist and A. Klug, ibid. 342, 825 (1989); J. R. Williamson, M. K. Raghuraman, T. R. Cech, Cell 59, 871 (1989); D. Sen and W Gilbert, Nature 344, 410 (1990); Z LIU, J D. Frantz, W. Gilbert, B Tye, Proc. Natl. Acad. Sci. U.S.A. 90, 3157 (1993); K. J. Woodford, R. M. Howell, K Usdin, J. Biol Chem. 269, 27029 (1994); M. Fry and L. A. Loeb, Proc. Natl. Acad Sci. U.S A 91, 4950 (1994)
41. J S. Lee, G. D. Burkhoider, L. J. P Latimer, B. L. Haug, R. P. Braun, Nucleic Acids Res. 15, 1047 (1987); S. M Mirkin and M D Frank-Kamenetski, Annu. Rev Biomol Struct 23, 541 (1994), M. J. Ulrich, W. J. Gray, T. J. Ley, J. Biol Chem. 267, 18649 (1992), Y. Kohwi and T. Kohwi-Shigematsu, Proc. Natl. Acad Sci. U.S.A. 85, 3781 (1988); A J. Kinniburgh, Nucleic Acids Res 17, 7771 (1989); D Sen and W. Gilbert, Nature 334, 364 (1988), W I Sundquist and A. Klug, ibid. 342, 825 (1989); J. R. Williamson, M. K. Raghuraman, T. R. Cech, Cell 59, 871 (1989); D. Sen and W Gilbert, Nature 344, 410 (1990); Z LIU, J D. Frantz, W. Gilbert, B Tye, Proc. Natl. Acad. Sci. U.S.A. 90, 3157 (1993); K. J. Woodford, R. M. Howell, K Usdin, J. Biol Chem. 269, 27029 (1994); M. Fry and L. A. Loeb, Proc. Natl. Acad Sci. U.S A 91, 4950 (1994)
42. J S. Lee, G. D. Burkhoider, L. J. P Latimer, B. L. Haug, R. P. Braun, Nucleic Acids Res. 15, 1047 (1987); S. M Mirkin and M D Frank-Kamenetski, Annu. Rev Biomol Struct 23, 541 (1994), M. J. Ulrich, W. J. Gray, T. J. Ley, J. Biol Chem. 267, 18649 (1992), Y. Kohwi and T. Kohwi-Shigematsu, Proc. Natl. Acad Sci. U.S.A. 85, 3781 (1988); A J. Kinniburgh, Nucleic Acids Res 17, 7771 (1989); D Sen and W. Gilbert, Nature 334, 364 (1988), W I Sundquist and A. Klug, ibid. 342, 825 (1989); J. R. Williamson, M. K. Raghuraman, T. R. Cech, Cell 59, 871 (1989); D. Sen and W Gilbert, Nature 344, 410 (1990); Z LIU, J D. Frantz, W. Gilbert, B Tye, Proc. Natl. Acad. Sci. U.S.A. 90, 3157 (1993); K. J. Woodford, R. M. Howell, K Usdin, J. Biol Chem. 269, 27029 (1994); M. Fry and L. A. Loeb, Proc. Natl. Acad Sci. U.S A 91, 4950 (1994)
43. J S. Lee, G. D. Burkhoider, L. J. P Latimer, B. L. Haug, R. P. Braun, Nucleic Acids Res. 15, 1047 (1987); S. M Mirkin and M D Frank-Kamenetski, Annu. Rev Biomol Struct 23, 541 (1994), M. J. Ulrich, W. J. Gray, T. J. Ley, J. Biol Chem. 267, 18649 (1992), Y. Kohwi and T. Kohwi-Shigematsu, Proc. Natl. Acad Sci. U.S.A. 85, 3781 (1988); A J. Kinniburgh, Nucleic Acids Res 17, 7771 (1989); D Sen and W. Gilbert, Nature 334, 364 (1988), W I Sundquist and A. Klug, ibid. 342, 825 (1989); J. R. Williamson, M. K. Raghuraman, T. R. Cech, Cell 59, 871 (1989); D. Sen and W Gilbert, Nature 344, 410 (1990); Z LIU, J D. Frantz, W. Gilbert, B Tye, Proc. Natl. Acad. Sci. U.S.A. 90, 3157 (1993); K. J. Woodford, R. M. Howell, K Usdin, J. Biol Chem. 269, 27029 (1994); M. Fry and L. A. Loeb, Proc. Natl. Acad Sci. U.S A 91, 4950 (1994)
44. J S. Lee, G. D. Burkhoider, L. J. P Latimer, B. L. Haug, R. P. Braun, Nucleic Acids Res. 15, 1047 (1987); S. M Mirkin and M D Frank-Kamenetski, Annu. Rev Biomol Struct 23, 541 (1994), M. J. Ulrich, W. J. Gray, T. J. Ley, J. Biol Chem. 267, 18649 (1992), Y. Kohwi and T. Kohwi-Shigematsu, Proc. Natl. Acad Sci. U.S.A. 85, 3781 (1988); A J. Kinniburgh, Nucleic Acids Res 17, 7771 (1989); D Sen and W. Gilbert, Nature 334, 364 (1988), W I Sundquist and A. Klug, ibid. 342, 825 (1989); J. R. Williamson, M. K. Raghuraman, T. R. Cech, Cell 59, 871 (1989); D. Sen and W Gilbert, Nature 344, 410 (1990); Z LIU, J D. Frantz, W. Gilbert, B Tye, Proc. Natl. Acad. Sci. U.S.A. 90, 3157 (1993); K. J. Woodford, R. M. Howell, K Usdin, J. Biol Chem. 269, 27029 (1994); M. Fry and L. A. Loeb, Proc. Natl. Acad Sci. U.S A 91, 4950 (1994)
45. J S. Lee, G. D. Burkhoider, L. J. P Latimer, B. L. Haug, R. P. Braun, Nucleic Acids Res. 15, 1047 (1987); S. M Mirkin and M D Frank-Kamenetski, Annu. Rev Biomol Struct 23, 541 (1994), M. J. Ulrich, W. J. Gray, T. J. Ley, J. Biol Chem. 267, 18649 (1992), Y. Kohwi and T. Kohwi-Shigematsu, Proc. Natl. Acad Sci. U.S.A. 85, 3781 (1988); A J. Kinniburgh, Nucleic Acids Res 17, 7771 (1989); D Sen and W. Gilbert, Nature 334, 364 (1988), W I Sundquist and A. Klug, ibid. 342, 825 (1989); J. R. Williamson, M. K. Raghuraman, T. R. Cech, Cell 59, 871 (1989); D. Sen and W Gilbert, Nature 344, 410 (1990); Z LIU, J D. Frantz, W. Gilbert, B Tye, Proc. Natl. Acad. Sci. U.S.A. 90, 3157 (1993); K. J. Woodford, R. M. Howell, K Usdin, J. Biol Chem. 269, 27029 (1994); M. Fry and L. A. Loeb, Proc. Natl. Acad Sci. U.S A 91, 4950 (1994)
46. J S. Lee, G. D. Burkhoider, L. J. P Latimer, B. L. Haug, R. P. Braun, Nucleic Acids Res. 15, 1047 (1987); S. M Mirkin and M D Frank-Kamenetski, Annu. Rev Biomol Struct 23, 541 (1994), M. J. Ulrich, W. J. Gray, T. J. Ley, J. Biol Chem. 267, 18649 (1992), Y. Kohwi and T. Kohwi-Shigematsu, Proc. Natl. Acad Sci. U.S.A. 85, 3781 (1988); A J. Kinniburgh, Nucleic Acids Res 17, 7771 (1989); D Sen and W. Gilbert, Nature 334, 364 (1988), W I Sundquist and A. Klug, ibid. 342, 825 (1989); J. R. Williamson, M. K. Raghuraman, T. R. Cech, Cell 59, 871 (1989); D. Sen and W Gilbert, Nature 344, 410 (1990); Z LIU, J D. Frantz, W. Gilbert, B Tye, Proc. Natl. Acad. Sci. U.S.A. 90, 3157 (1993); K. J. Woodford, R. M. Howell, K Usdin, J. Biol Chem. 269, 27029 (1994); M. Fry and L. A. Loeb, Proc. Natl. Acad Sci. U.S A 91, 4950 (1994)
47. J S. Lee, G. D. Burkhoider, L. J. P Latimer, B. L. Haug, R. P. Braun, Nucleic Acids Res. 15, 1047 (1987); S. M Mirkin and M D Frank-Kamenetski, Annu. Rev Biomol Struct 23, 541 (1994), M. J. Ulrich, W. J. Gray, T. J. Ley, J. Biol Chem. 267, 18649 (1992), Y. Kohwi and T. Kohwi-Shigematsu, Proc. Natl. Acad Sci. U.S.A. 85, 3781 (1988); A J. Kinniburgh, Nucleic Acids Res 17, 7771 (1989); D Sen and W. Gilbert, Nature 334, 364 (1988), W I Sundquist and A. Klug, ibid. 342, 825 (1989); J. R. Williamson, M. K. Raghuraman, T. R. Cech, Cell 59, 871 (1989); D. Sen and W Gilbert, Nature 344, 410 (1990); Z LIU, J D. Frantz, W. Gilbert, B Tye, Proc. Natl. Acad. Sci. U.S.A. 90, 3157 (1993); K. J. Woodford, R. M. Howell, K Usdin, J. Biol Chem. 269, 27029 (1994); M. Fry and L. A. Loeb, Proc. Natl. Acad Sci. U.S A 91, 4950 (1994)
48. J S. Lee, G. D. Burkhoider, L. J. P Latimer, B. L. Haug, R. P. Braun, Nucleic Acids Res. 15, 1047 (1987); S. M Mirkin and M D Frank-Kamenetski, Annu. Rev Biomol Struct 23, 541 (1994), M. J. Ulrich, W. J. Gray, T. J. Ley, J. Biol Chem. 267, 18649 (1992), Y. Kohwi and T. Kohwi-Shigematsu, Proc. Natl. Acad Sci. U.S.A. 85, 3781 (1988); A J. Kinniburgh, Nucleic Acids Res 17, 7771 (1989); D Sen and W. Gilbert, Nature 334, 364 (1988), W I Sundquist and A. Klug, ibid. 342, 825 (1989); J. R. Williamson, M. K. Raghuraman, T. R. Cech, Cell 59, 871 (1989); D. Sen and W Gilbert, Nature 344, 410 (1990); Z LIU, J D. Frantz, W. Gilbert, B Tye, Proc. Natl. Acad. Sci. U.S.A. 90, 3157 (1993); K. J. Woodford, R. M. Howell, K Usdin, J. Biol Chem. 269, 27029 (1994); M. Fry and L. A. Loeb, Proc. Natl. Acad Sci. U.S A 91, 4950 (1994)
49. J S. Lee, G. D. Burkhoider, L. J. P Latimer, B. L. Haug, R. P. Braun, Nucleic Acids Res. 15, 1047 (1987); S. M Mirkin and M D Frank-Kamenetski, Annu. Rev Biomol Struct 23, 541 (1994), M. J. Ulrich, W. J. Gray, T. J. Ley, J. Biol Chem. 267, 18649 (1992), Y. Kohwi and T. Kohwi-Shigematsu, Proc. Natl. Acad Sci. U.S.A. 85, 3781 (1988); A J. Kinniburgh, Nucleic Acids Res 17, 7771 (1989); D Sen and W. Gilbert, Nature 334, 364 (1988), W I Sundquist and A. Klug, ibid. 342, 825 (1989); J. R. Williamson, M. K. Raghuraman, T. R. Cech, Cell 59, 871 (1989); D. Sen and W Gilbert, Nature 344, 410 (1990); Z LIU, J D. Frantz, W. Gilbert, B Tye, Proc. Natl. Acad. Sci. U.S.A. 90, 3157 (1993); K. J. Woodford, R. M. Howell, K Usdin, J. Biol Chem. 269, 27029 (1994); M. Fry and L. A. Loeb, Proc. Natl. Acad Sci. U.S A 91, 4950 (1994)
50. J S. Lee, G. D. Burkhoider, L. J. P Latimer, B. L. Haug, R. P. Braun, Nucleic Acids Res. 15, 1047 (1987); S. M Mirkin and M D Frank-Kamenetski, Annu. Rev Biomol Struct 23, 541 (1994), M. J. Ulrich, W. J. Gray, T. J. Ley, J. Biol Chem. 267, 18649 (1992), Y. Kohwi and T. Kohwi-Shigematsu, Proc. Natl. Acad Sci. U.S.A. 85, 3781 (1988); A J. Kinniburgh, Nucleic Acids Res 17, 7771 (1989); D Sen and W. Gilbert, Nature 334, 364 (1988), W I Sundquist and A. Klug, ibid. 342, 825 (1989); J. R. Williamson, M. K. Raghuraman, T. R. Cech, Cell 59, 871 (1989); D. Sen and W Gilbert, Nature 344, 410 (1990); Z LIU, J D. Frantz, W. Gilbert, B Tye, Proc. Natl. Acad. Sci. U.S.A. 90, 3157 (1993); K. J. Woodford, R. M. Howell, K Usdin, J. Biol Chem. 269, 27029 (1994); M. Fry and L. A. Loeb, Proc. Natl. Acad Sci. U.S A 91, 4950 (1994)
51. J S. Lee, G. D. Burkhoider, L. J. P Latimer, B. L. Haug, R. P. Braun, Nucleic Acids Res. 15, 1047 (1987); S. M Mirkin and M D Frank-Kamenetski, Annu. Rev Biomol Struct 23, 541 (1994), M. J. Ulrich, W. J. Gray, T. J. Ley, J. Biol Chem. 267, 18649 (1992), Y. Kohwi and T. Kohwi-Shigematsu, Proc. Natl. Acad Sci. U.S.A. 85, 3781 (1988); A J. Kinniburgh, Nucleic Acids Res 17, 7771 (1989); D Sen and W. Gilbert, Nature 334, 364 (1988), W I Sundquist and A. Klug, ibid. 342, 825 (1989); J. R. Williamson, M. K. Raghuraman, T. R. Cech, Cell 59, 871 (1989); D. Sen and W Gilbert, Nature 344, 410 (1990); Z LIU, J D. Frantz, W. Gilbert, B Tye, Proc. Natl. Acad. Sci. U.S.A. 90, 3157 (1993); K. J. Woodford, R. M. Howell, K Usdin, J. Biol Chem. 269, 27029 (1994); M. Fry and L. A. Loeb, Proc. Natl. Acad Sci. U.S A 91, 4950 (1994)
52. Third-strand binding was measured with a gel mobility-shift assay as described [R. H Durland et al., Biochemistry 30, 9246 (1991)].
53. We thank J. H J. Hoeijmakers for providing the pSLME6(+) vector and K. Kraemer and D D. Levy for providing the XPA cell lines. We also thank P. A. Havre, T Yeasky, F. P. Gasparro, L Cabral, S. J. Baserga, and R. Franklin for their help. Supported by the Charles E. Culpeper Foundation, the Leukemia Society of America, the American Cancer Society (grant CN-128), and NIH (grants ES05775 and CA64186).