1QFQ

Bacteriophage Lambda N-protein-NutboxB-RNA Complex

Summary for 1QFQ

• Entry DOI: 10.2210/pdb1qfq/pdb
• Descriptor: 15-mer nutRboxB RNA hairpin, 36-mer N-terminal peptide of the N protein (2 entities in total)
• Functional Keywords: bacteriophage lambda; antitermination; peptide-rna-complex; n-nut; gnra tetraloop; bent-alpha-helix; peptide-rna-recognition, transcription-rna complex, transcription/rna
• Biological source: Enterobacteria phage lambda; More
• Total number of polymer chains: 2
• Total formula weight: 8861.58

Authors

Schaerpf, M.,Sticht, H.,Roesch, P. (deposition date: 1999-04-12, release date: 1999-04-20, Last modification date: 2023-12-27)

Primary citation

Schaerpf, M.,Sticht, H.,Schweimer, K.,Boehm, M.,Hoffmann, S.,Roesch, P.
Antitermination in bacteriophage lambda. The structure of the N36 peptide-boxB RNA complex
Eur.J.Biochem., 267:2397-2408, 2000

PubMed Abstract: The solution structure of a 15-mer nutRboxB RNA hairpin complexed with the 36-mer N-terminal peptide of the N protein (N36) from bacteriophage lambda was determined by 2D and 3D homonuclear and heteronuclear magnetic resonance spectroscopy. These 36 amino acids include the arginine-rich motif of the N protein involved in transcriptional antitermination of phage lambda. Upon complex formation with boxB RNA, the synthetic N36 peptide binds tightly to the major groove of the boxB hairpin through hydrophobic and electrostatic interactions forming a bent alpha helix. Four nucleotides of the GAAAA pentaloop of the boxB RNA adopt a GNRA-like tetraloop fold in the complex. The formation of a GAAA tetraloop involves a loop-closing sheared base pair (G6-A10), base stacking of three adenines (A7, A8, and A10), and extrusion of one nucleotide (A9) from the loop, as observed previously for the complex of N(1-22) peptide and the nutLboxB RNA [Legault, P., Li, J., Mogridge, J., Kay, L.E. & Greenblatt, J. (1998) Cell 93, 289-299]. Stacking of the bases is extended by the indole-ring of Trp18 which also forms hydrophobic contacts to the side-chains of Leu24, Leu25, and Val26. Based on the structure of the complex, three mutant peptides were synthesized and investigated by CD and NMR spectroscopy in order to determine the role of particular residues for complex formation. These studies revealed very distinct amino-acid requirements at positions 3, 4, and 8, while replacement of Trp18 with tyrosine did not result in any gross structural changes.

PubMed: 10759866
DOI: 10.1046/j.1432-1327.2000.01251.x

Experimental method

SOLUTION NMR