Summary
The first three zinc fingers (ZF1-3) of transcription factor IIIA (TFIIIA) from Xenopus have been shown to contribute the majority of the binding energy to the intact TFIIIA-DNA interaction [Liao et al. (1992) J. Mol. Biol., 223, 857–871]. We have expressed a 92-amino acid polypeptide containing the three N-terminal zinc fingers of TFIIIA. This three-fingered polypeptide has been isotopically labeled with 15N and 13C in E. coli and purified to homogeneity. Assignment of backbone 1H, 15N, aliphatic 1H and 13C and aromatic 1H and 13C resonances of ΔNZF1-3 has been obtained using a combination of single-, double-and triple-resonance multidimensional NMR experiments. The secondary structures for each finger have been determined from NOE connectivities, 3JNHα values and chemical shifts. The results show that each finger folds into a canonical β-sheet-helix zinc finger structural motif, while the linkers adopt an extended structure. The helix between the two histidine ligands in ZF3 is distorted by zinc coordination, to accommodate the presence of four intervening amino acids instead of three as in ZF1 and ZF2.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bax, A. and Subramanian, S.J. (1986) J. Magn. Reson., 67, 565–569.
Bax, A. (1989) Annu. Rev. Biochem., 58, 233–256.
Bax, A., Ikura, M., Kay, L.E., Torchia, D. and Tschudin, R. (1990a) J. Magn. Reson., 86, 304–318.
Bax, A., Clore, G.M. and Gronenborn, A.M. (1990b) J. Magn. Reson., 88, 425–431.
Bodenhausen, G. and Ruben, D.J. (1980) Chem. Phys. Lett., 69, 185–187.
Braunschweiler, L. and Ernst, R.R. (1983) J. Magn. Reson., 53, 521–526.
Braunschweiler, L., Bodenhausen, G. and Ernst, R.R. (1984) Mol. Phys., 48, 535–560.
Bundi, A. and Wüthrich, K. (1979) Biopolymers, 18, 285–297.
Cavanagh, J. and Rance, M. (1990) J. Magn. Reson., 88, 72–76.
Clemens, K., Liao, X., Wolf, V., Wright, P.E. and Gottesfeld, J. (1992) Proc. Natl. Acad. Sci. USA, 89, 10822–10826.
Clemens, K., Wolf, V., McBryant, S.J., Zhang, P., Liao, X., Wright, P.E. and Gottesfeld, J. (1993) Science, 260, 530–533.
Clore, G.M. and Gronenborn, A.M. (1989) Crit. Rev. Biochem. Mol. Biol., 24, 479–564.
Clore, G.M., Kay, L.E., Bax, A. and Gronenborn, A.M. (1991) Biochemistry, 30, 12–18.
El-Baradi, T. and Pieler, T. (1991) Mech. Dev., 35, 155–169.
Engelke, D.R., Ng, S.-Y., Shastry, B.S. and Roeder, D.G. (1980) Cell, 19, 717–728.
Fesik, S.W. and Zuiderweg, E.R.P. (1990) Q. Rev. Biophys., 23, 97–131.
Ginsberg, A.M., King, B.O. and Roeder, R.G. (1984) Cell, 39, 479–489.
Honda, B.M. and Roeder, R.G. (1980) Cell, 22, 119–126.
Howarth, O.W. and Lilley, D.M.J. (1978) Prog. NMR Spectrosc., 12, 1–40.
Ikura, M., Kay, L.E. and Bax, A. (1991) J. Biomol. NMR, 1, 299–304.
Jeener, J., Meier, B.H., Bachmann, P. and Ernst, R.R. (1979) J. Chem. Phys., 71, 4546–4553.
Klevit, R.E., Herriott, J.R. and Horvath, S. (1991) Proteins, 7, 214–226.
Kochoyan, M., Havel, T.F., Nguyen, D.T., Dahl, C.E., Keutmann, H.T. and Weiss, M.A. (1991a) Biochemistry, 30, 3371–3386.
Kochoyan, M., Keutmann, H.T. and Weiss, M.A. (1991b) Biochemistry, 30, 7063–7072.
Lee, M.S., Gippert, G.P., Soman, K.V., Case, D.A. and Wright, P.E. (1989) Science, 245, 635–637.
Lee, M.S., PalmerIII, A.G. and Wright, P.E. (1992a) J. Biomol. NMR, 2, 307–322.
Lee, M.S., Mortishire-Smith, R.J. and Wright, P.E. (1992b) FEBS Lett., 309, 29–32.
Liao, X., Clemens, K.R., Tennant, L., Wright, P.E. and Gottesfeld, J.M. (1992) J. Mol. Biol., 223, 857–871.
Live, D.H., Davis, D.G., Agosta, W.C. and Cowburn, D. (1984) J. Am. Chem. Soc., 106, 1939–1941.
Marion, D. and Wüthrich, K. (1983) Biochem. Biophys. Res. Commun., 113, 967–974.
Marion, D., Ikura, M., Tschudin, R. and Bax, A. (1989) J. Magn. Reson., 85, 393–399.
McIntosh, L.P. and Dahlquist, F.W. (1990) Q. Rev. Biophys., 23, 1–38.
Messerle, B.A., Wider, G., Otting, G., Weber, C. and Wüthrich, K. (1989) J. Magn. Reson., 85, 608–613.
Miller, J., McLachlan, A.D. and Klug, A. (1985) EMBO J., 4, 1609–1614.
Morris, G.A. and Freeman, R. (1979) J. Am. Chem. Soc., 101, 760–762.
Neri, D., Otting, G. and Wüthrich, K. (1990) J. Am. Chem. Soc., 112, 3663–3665.
Norwood, T.J., Boyd, J., Heritage, J.E., Soffe, N. and Campbell, I.D. (1990) J. Magn. Reson., 87, 488–501.
Omichinski, J.G., Clore, G.M., Appella, E., Sakaguchi, K. and Gronenborn, A.M. (1990) Biochemistry, 29, 9324–9334.
Omichinski, J.G., Clore, G.M., Robien, M., Sakaguchi, K., Appella, E. and Gronenborn, A.M. (1992) Biochemistry, 31, 3907–3917.
PalmerIII, A.G., Cavanagh, J., Byrd, R.A. and Rance, M. (1991) J. Magn. Reson., 96, 416–424.
PalmerIII, A.G., Fairbrother, W.J., Cavanagh, J., Wright, P.E. and Rance, M. (1992) J. Biomol. NMR, 2, 103–108.
Pavletich, N.P. and Pabo, C.O. (1991) Science, 252, 809–917.
Pelham, H.R. and Brown, D.D. (1980) Proc. Natl. Acad. Sci. USA, 77, 4170–4174.
Richarz, R. and Wüthrich, K. (1970) Biopolymers, 17, 2133–2141.
Rance, M. and Wright, P.E. (1986) J. Magn. Reson., 66, 372–378.
Sakonju, S., Bogenhagen, D.F. and Brown, D.D. (1980) Cell, 19, 13–25.
Shaka, A.J., Barker, P.B. and Freeman, R. (1985) J. Magn. Reson., 64, 547–552.
Shaka, A.J., Lee, C.J. and Pines, A. (1988) J. Magn. Reson., 77, 274–293.
Smith, D.R., Jackson, I.J. and Brown, D.D. (1984) Cell, 37, 645–652.
Spera, A.J. and Bax, A. (1991) J. Am. Chem. Soc., 113, 5490–5492.
Vrana, K.E., Churchill, M.E.A., Tullius, T.D. and Brown, D.D. (1988) Mol. Cell. Biol., 8, 1684–1696.
Wishart, D.S., Sykes, B.D. and Richards, F.M. (1991) J. Mol. Biol., 222, 311–333.
Wüthrich, K. (1986) NMR of Proteins and Nucleic Acids, Wiley, New York, NY.
Wüthrich, K. (1989) Acc. Chem., 22, 36–44.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Liao, X., Clemens, K., Cavanagh, J. et al. 1H, 15N and 13C resonance assignments for the first three zinc fingers of transcription factor IIIA. J Biomol NMR 4, 433–454 (1994). https://doi.org/10.1007/BF00179350
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00179350