Tat-SF1: Cofactor for stimulation of transcriptional elongation by HIV- 1 Tat

Science

Volumn 274, Issue 5287, 1996, Pages 605-610

  Zhou, Qiang ^a   Sharp, Phillip A ^a  

^a MASSACHUSETTS INSTITUTE OF TECHNOLOGY   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

COMPLEMENTARY DNA; RNA POLYMERASE II; TRANSACTIVATOR PROTEIN; TRANSCRIPTION FACTOR;

ARTICLE; DNA SEQUENCE; GENE EXPRESSION REGULATION; HELA CELL; HUMAN; HUMAN CELL; HUMAN IMMUNODEFICIENCY VIRUS 1; NONHUMAN; PRIORITY JOURNAL; SARCOMA; TRANSACTIVATION; TRANSCRIPTION INITIATION; VIRUS CELL INTERACTION; VIRUS REPLICATION;

AMINO ACID SEQUENCE; DNA, COMPLEMENTARY; ELECTROPHORESIS, POLYACRYLAMIDE GEL; GENE EXPRESSION; GENE PRODUCTS, TAT; HELA CELLS; HETEROGENEOUS-NUCLEAR RIBONUCLEOPROTEINS; HIV LONG TERMINAL REPEAT; HIV-1; HUMANS; IMMUNOBLOTTING; MOLECULAR SEQUENCE DATA; MOLECULAR WEIGHT; NEOPLASM PROTEINS; PHOSPHORYLATION; PROMOTER REGIONS (GENETICS); PROTEIN KINASES; RIBONUCLEOPROTEINS; RNA, VIRAL; RNA-BINDING PROTEIN EWS; RNA-BINDING PROTEIN FUS; SEQUENCE HOMOLOGY, AMINO ACID; TRANS-ACTIVATION (GENETICS); TRANS-ACTIVATORS; TRANSFECTION;

HUMAN IMMUNODEFICIENCY VIRUS; HUMAN IMMUNODEFICIENCY VIRUS 1;

EID: 0029834383     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.274.5287.605     Document Type: Article

References (49)

1. W. S. Blair, R. A. Fridell, B. R. Cullen, EMBO J. 15, 101 (1996); J. Blau et al., Mol. Cell. Biol. 16, 2044 (1996); K. Yankulov, J. Blau, T. Purton, S. Roberts, D. L. Bentley, Cell 77, 749 (1994).
2. W. S. Blair, R. A. Fridell, B. R. Cullen, EMBO J. 15, 101 (1996); J. Blau et al., Mol. Cell. Biol. 16, 2044 (1996); K. Yankulov, J. Blau, T. Purton, S. Roberts, D. L. Bentley, Cell 77, 749 (1994).
3. W. S. Blair, R. A. Fridell, B. R. Cullen, EMBO J. 15, 101 (1996); J. Blau et al., Mol. Cell. Biol. 16, 2044 (1996); K. Yankulov, J. Blau, T. Purton, S. Roberts, D. L. Bentley, Cell 77, 749 (1994).
4. B. R. Cullen, Infect. Agents Dis. 3, 68 (1994); A. D. Frankel, Curr. Opin. Genet. Dev. 2, 293 (1992); K. A. Jones and B. M. Peterlin, Annu. Rev. Biochem. 63, 717 (1994); J. Kam and M. A. Graeble, Trends Genet. 8, 365 (1992); R. B. Gaynor, Curr. Top. Microbiol. Immunol. 193, 51 (1995).
5. B. R. Cullen, Infect. Agents Dis. 3, 68 (1994); A. D. Frankel, Curr. Opin. Genet. Dev. 2, 293 (1992); K. A. Jones and B. M. Peterlin, Annu. Rev. Biochem. 63, 717 (1994); J. Kam and M. A. Graeble, Trends Genet. 8, 365 (1992); R. B. Gaynor, Curr. Top. Microbiol. Immunol. 193, 51 (1995).
6. B. R. Cullen, Infect. Agents Dis. 3, 68 (1994); A. D. Frankel, Curr. Opin. Genet. Dev. 2, 293 (1992); K. A. Jones and B. M. Peterlin, Annu. Rev. Biochem. 63, 717 (1994); J. Kam and M. A. Graeble, Trends Genet. 8, 365 (1992); R. B. Gaynor, Curr. Top. Microbiol. Immunol. 193, 51 (1995).
7. B. R. Cullen, Infect. Agents Dis. 3, 68 (1994); A. D. Frankel, Curr. Opin. Genet. Dev. 2, 293 (1992); K. A. Jones and B. M. Peterlin, Annu. Rev. Biochem. 63, 717 (1994); J. Kam and M. A. Graeble, Trends Genet. 8, 365 (1992); R. B. Gaynor, Curr. Top. Microbiol. Immunol. 193, 51 (1995).
8. B. R. Cullen, Infect. Agents Dis. 3, 68 (1994); A. D. Frankel, Curr. Opin. Genet. Dev. 2, 293 (1992); K. A. Jones and B. M. Peterlin, Annu. Rev. Biochem. 63, 717 (1994); J. Kam and M. A. Graeble, Trends Genet. 8, 365 (1992); R. B. Gaynor, Curr. Top. Microbiol. Immunol. 193, 51 (1995).
9. S. Feng and E. C. Holland, Nature 334, 165 (1988); J. A. Garcia et al., EMBO J. 8, 765 (1989); M. J. Selby, E. S. Bain, P. A. Luciw, B. M. Peterlin, Genes Dev. 3, 547 (1989).
10. S. Feng and E. C. Holland, Nature 334, 165 (1988); J. A. Garcia et al., EMBO J. 8, 765 (1989); M. J. Selby, E. S. Bain, P. A. Luciw, B. M. Peterlin, Genes Dev. 3, 547 (1989).
11. S. Feng and E. C. Holland, Nature 334, 165 (1988); J. A. Garcia et al., EMBO J. 8, 765 (1989); M. J. Selby, E. S. Bain, P. A. Luciw, B. M. Peterlin, Genes Dev. 3, 547 (1989).
12. A. Kibel, O. Iliopoulos, J. A. De Caprio, W. G. Kaelin Jr., Science 269, 1444 (1995).
13. D. R. Duan et al., ibid., p. 1402.
14. T. Aso, W. S. Lane, J. W. Conaway, R. C. Conaway, ibid., p. 1439.
15. Q. Zhou and P. A. Sharp, EMBO J. 14, 321 (1995).
16. _, unpublished data.
17. HeLa nuclear extract in buffer D-0.1 M KCI [20 mM Hepes-KOH (pH 7.9), 20% (v/v) glycerol, 0.1 M KCI, 0.2 mM EDTA, 0.5 mM dithiothreitol, 0.5 mM phenylmethylsulfonyl fluoride] was loaded onto a phosphocellulose column preequilibrated with the same buffer. The flow-through was loaded onto a DEAE-Sepharose FF (Pharmacia) matrix column preequilibrated with buffer D-0.1 M KCI. After the column was washed with the same buffer, Tat-SF activity was eluted from the column with buffer D-0.3 M KCI. This fraction was dialyzed against buffer D-0.1 M KCI and applied to a Q-Sepharose FF (Pharmacia) matrix column preequilibrated with the same buffer. The column was washed with buffer D-0.1 M KCI, and the bound proteins were eluted with a gradient of 0.1 to 0.7 M KCI in buffer D. Fractions were analyzed for Tat-SF activity in reconstituted transcription assays and for pp140 in kinase reactions. The 0.4 to 0.5 M KCI Q-Sepharose fraction containing Tat-SF activity and pp 140 was dialyzed against buffer D-0.1 M KCI and applied to a heparin Sepharose column. After the column was washed extensively with buffer D-0.1 M KCI, Tat-SF/pp140 was eluted with in-creasing salt concentrations and was detected mostly in 0.2 to 0.4 M KCI fractions. These fractions were pooled, dialyzed against buffer D-0.1 M KCI, and loaded onto a glutathione Sepharose (Pharmacia) column containing GST-Tat fusion proteins. After the column was washed with buffer D-0.4 M KCI, Tat-SF/pp 140 was eluted from the column with buffer D containing 1.4 M KCI. The estimated overall purification after these steps was -3000-fold. In the experiment shown in Fig. 3, the 0.2 to 0.4 M KCI heparin Sepharose fraction containing Tat-SF activity was subjected to fractionation through an Affi-Gel 10 matrix column (Bio-Rad) containing immobilized Tat. Tat-SF activity was eluted from the column with increasing salt concentrations. The 0.6 M KCI fraction was analyzed as described in Fig 3.
18. T. O'Brien, S, Hardin, A. Greenleaf, J. T. Lis, Nature 370, 75 (1994); M. E. Dahmus, Biochim. Biophys. Acta. 1261, 171 (1995).
19. T. O'Brien, S, Hardin, A. Greenleaf, J. T. Lis, Nature 370, 75 (1994); M. E. Dahmus, Biochim. Biophys. Acta. 1261, 171 (1995).
20. A. P. Rice and F. Carlotti, J. Virol. 64, 1864 (1990).
21. The Tat-SF/pp140 fraction eluted from the GST-Tat column was subjected to SDS-polyacrylamide gel electrophoresis (PAGE), and the pp 140 polypeptide was blotted onto a nitrocellulose membrane. Approximately 15 μg of pp140 were recovered from the membrane and subjected to digestion with lys-C. Six major peptides were obtained and microsequenced. One of the peptides (KMNAQETATGMAFEEPIDE) was contained in the sequence of EST60354 in the Washington University-Merck EST database. An Xho l-Eco Rl fragment corresponding to the COOH-terminus of the Tat-SF1 gene and its 3′ untranslated region was labeled and used as a probe to screen a λZipLox (Gibco BRL) cDNA library prepared from human HL60 cells. Complementary DNAs were recovered from seven independent plaques in the autonomously replicating plasmid pZL1 as instructed by the manufacturer (Gibco BRL). The largest cDNA clone containing the full-length Tat-SF1 gene was named pZL-Tat-SF1-4b and was sequenced by dideoxy-DNA sequencing with T7 DNA polymerase.
22. D. R. Marshak and D. Carroll, Methods Enzymol. 200, 134 (1991).
23. D. J. Kenan, C. C. Query, J. D. Keene, Trends Biochem. Sci. 16, 214 (1991).
24. O. Delattre et al., Nature 359, 162 (1992); P. H. Sorensen et al., Nature Genet. 6, 146 (1994).
25. O. Delattre et al., Nature 359, 162 (1992); P. H. Sorensen et al., Nature Genet. 6, 146 (1994).
26. A. Crozat, P. Aman, N. Mandahl, D. Ron, Nature 363, 640 (1993); T. H. Rabbitts, A. Forster, R. Larson, P. Nathan, Nature Genet. 4, 175 (1993).
27. A. Crozat, P. Aman, N. Mandahl, D. Ron, Nature 363, 640 (1993); T. H. Rabbitts, A. Forster, R. Larson, P. Nathan, Nature Genet. 4, 175 (1993).
28. M. Ladanyi, Diagn. Mot. Pathol. 4, 162 (1995); T. H. Rabbitts, Nature 372, 143 (1994).
29. M. Ladanyi, Diagn. Mot. Pathol. 4, 162 (1995); T. H. Rabbitts, Nature 372, 143 (1994).
30. S. E. Harper, Y. Qiu, P. A. Sharp, Proc. Natl. Acad. Sci. U.S.A. 93, 8536 (1996).
31. J. W. Lillie and M. R. Green, Nature 338, 39 (1989).
32. H. Kato et al., Genes Dev. 6, 655 (1992); R. A. Marciniak and P. A. Sharp, EMBO J. 10, 4189 (1991).
33. H. Kato et al., Genes Dev. 6, 655 (1992); R. A. Marciniak and P. A. Sharp, EMBO J. 10, 4189 (1991).
34. M. G. Izban and D. S Luse, Genes Dev. 6, 1342 (1992); D. Wang and D. K. Hawley, Proc, Natl. Acad. Sci. U.S.A. 90, 843 (1993)
35. M. G. Izban and D. S Luse, Genes Dev. 6, 1342 (1992); D. Wang and D. K. Hawley, Proc, Natl. Acad. Sci. U.S.A. 90, 843 (1993)
36. E. Bengal, O. Flores, A. Krauskopf, D. Reinberg, Y. Aloni, Mol. Cell. Biol. 11, 1195 (1991); J. Greenblatt, J. R. Nodwell, S. W. Mason, Nature 364, 401 (1993).
37. E. Bengal, O. Flores, A. Krauskopf, D. Reinberg, Y. Aloni, Mol. Cell. Biol. 11, 1195 (1991); J. Greenblatt, J. R. Nodwell, S. W. Mason, Nature 364, 401 (1993).
38. C. H. Herrmann and A. P. Rice, J. Virol. 69, 1612 (1995).
39. N. A. McMillan et al., Virology 213, 413 (1995).
40. W. A. May et al., Mol. Cell. Biol. 13, 7393 (1993); H. Zinszner, R. Albalat, D. Ron, Genes Dev. 8, 2513 (1994); D. D. Prasad, M. Ouchida, L. Lee, V. N. Rao, E. S. Reddy, Oncogene 9, 3717 (1994).
41. W. A. May et al., Mol. Cell. Biol. 13, 7393 (1993); H. Zinszner, R. Albalat, D. Ron, Genes Dev. 8, 2513 (1994); D. D. Prasad, M. Ouchida, L. Lee, V. N. Rao, E. S. Reddy, Oncogene 9, 3717 (1994).
42. W. A. May et al., Mol. Cell. Biol. 13, 7393 (1993); H. Zinszner, R. Albalat, D. Ron, Genes Dev. 8, 2513 (1994); D. D. Prasad, M. Ouchida, L. Lee, V. N. Rao, E. S. Reddy, Oncogene 9, 3717 (1994).
43. P. J. Mitchell and R. Tjian, Science 245, 371 (1989).
44. S. F. Altschul, W. Gish, W. Miller, E. W. Myers, D. J, Lipman, J. Mol. Biol. 215, 403 (1990).
45. M. A. Truett et al., DNA 4, 333 (1985).
46. H. E. Gendelman et al., Proc. Natl. Acad. Sci. U.S.A. 83, 9759 (1986).
47. L. S. Tiley, P. H. Brown, B. R. Cullen, Virology 178, 560 (1990).
48. J. R. Neumann, C. A. Morency, K. O. Russian, BioTechniques 5, 444 (1987).
49. We are grateful to B. Pepinsky and Biogen for providing pure HIV Tat protein and Tat mutant TatΔC; to J. Borrow [Massachusetts Institute of Technology (MIT ) Center for Cancer Research] for human cDNA libraries; and to R. Cook (MIT Biopolymers Laboratory) for peptide sequencing We thank K. Luo, J. Borrow, and H. Kawasaki for valuable advice and discussions; and B. Blencowe, K. Cepek, G. Jones, K. Luo, and C. Query for helpful comments on the manuscript. We also thank M. Siafaca for secretarial support. Supported by grants from the National Institutes of Health (GM34277 and AI32486) to P.A.S., and partially supported by a National Cancer Institute Center core grant (CA14051). Q.Z. was supported by a postdoctoral fellowship of The Jane Coffin Childs Memorial Fund for Medical Research.