The Influence of C-terminal Extension on the Structure of the "J-domain" in E. Coli DnaJ

Overview

Journal Protein Sci

Specialty Biochemistry

Date 1999 Apr 21

PMID 10210198

Citations 22

Authors

K Huang

J M Flanagan

J H Prestegard

Affiliations

Soon will be listed here.

Abstract

Two different recombinant constructs of the N-terminal domain in Escherichia coli DnaJ were uniformly labeled with nitrogen-15 and carbon-13. One, DnaJ(1-78), contains the complete "J-domain," and the other, DnaJ(1-104), contains both the "J-domain" and a conserved "G/F" extension at the C-terminus. The three-dimensional structures of these proteins have been determined by heteronuclear NMR experiments. In both proteins the "J-domain" adopts a compact structure consisting of a helix-turn-helix-loop-helix-turn-helix motif. In contrast, the "G/F" region in DnaJ(1-104) does not fold into a well-defined structure. Nevertheless, the "G/F" region has been found to have an effect on the packing of the helices in the "J-domain" in DnaJ(1-104). Particularly, the interhelical angles between Helix IV and other helices are significantly different in the two structures. In addition, there are some local conformational changes in the loop region connecting the two central helices. These structural differences in the "J-domain" in the presence of the "G/F" region may be related to the observation that DnaJ (1-78) is incapable of stimulating the ATPase activity of the molecular chaperone protein DnaK despite evidence that sites mediating the binding of DnaJ to DnaK are located in the 1-78 segment.

Citing Articles

Structural remodeling of ribosome associated Hsp40-Hsp70 chaperones during co-translational folding.

Chen Y, Tsai B, Li N, Gao N Nat Commun. 2022; 13(1):3410.

PMID: 35701497 PMC: 9197937. DOI: 10.1038/s41467-022-31127-4.

A History of Molecular Chaperone Structures in the Protein Data Bank.

Bascos N, Landry S Int J Mol Sci. 2019; 20(24).

PMID: 31817979 PMC: 6940948. DOI: 10.3390/ijms20246195.

Structural and functional analysis of the Hsp70/Hsp40 chaperone system.

Liu Q, Liang C, Zhou L Protein Sci. 2019; 29(2):378-390.

PMID: 31509306 PMC: 6954727. DOI: 10.1002/pro.3725.

The Complex Phosphorylation Patterns that Regulate the Activity of Hsp70 and Its Cochaperones.

Velasco L, Dublang L, Moro F, Muga A Int J Mol Sci. 2019; 20(17).

PMID: 31450862 PMC: 6747476. DOI: 10.3390/ijms20174122.

Broadening the functionality of a J-protein/Hsp70 molecular chaperone system.

Schilke B, Ciesielski S, Ziegelhoffer T, Kamiya E, Tonelli M, Lee W PLoS Genet. 2017; 13(10):e1007084.

PMID: 29084221 PMC: 5679652. DOI: 10.1371/journal.pgen.1007084.

References

Sunshine M, Feiss M, Stuart J, Yochem J . A new host gene (groPC) necessary for lambda DNA replication. Mol Gen Genet. 1977; 151(1):27-34. DOI: 10.1007/BF00446909. View

Rice L, Brunger A . Torsion angle dynamics: reduced variable conformational sampling enhances crystallographic structure refinement. Proteins. 1994; 19(4):277-90. DOI: 10.1002/prot.340190403. View

Wuthrich K, Billeter M, Braun W . Pseudo-structures for the 20 common amino acids for use in studies of protein conformations by measurements of intramolecular proton-proton distance constraints with nuclear magnetic resonance. J Mol Biol. 1983; 169(4):949-61. DOI: 10.1016/s0022-2836(83)80144-2. View

Ohki M, Tamura F, Nishimura S, Uchida H . Nucleotide sequence of the Escherichia coli dnaJ gene and purification of the gene product. J Biol Chem. 1986; 261(4):1778-81. View

Bardwell J, Tilly K, Craig E, King J, Zylicz M, Georgopoulos C . The nucleotide sequence of the Escherichia coli K12 dnaJ+ gene. A gene that encodes a heat shock protein. J Biol Chem. 1986; 261(4):1782-5. View

Richards F, Kundrot C . Identification of structural motifs from protein coordinate data: secondary structure and first-level supersecondary structure. Proteins. 1988; 3(2):71-84. DOI: 10.1002/prot.340030202. View

Straus D, WALTER W, Gross C . Escherichia coli heat shock gene mutants are defective in proteolysis. Genes Dev. 1988; 2(12B):1851-8. DOI: 10.1101/gad.2.12b.1851. View

Straus D, Walter W, Gross C . DnaK, DnaJ, and GrpE heat shock proteins negatively regulate heat shock gene expression by controlling the synthesis and stability of sigma 32. Genes Dev. 1990; 4(12A):2202-9. DOI: 10.1101/gad.4.12a.2202. View

Liberek K, Marszalek J, Ang D, Georgopoulos C, Zylicz M . Escherichia coli DnaJ and GrpE heat shock proteins jointly stimulate ATPase activity of DnaK. Proc Natl Acad Sci U S A. 1991; 88(7):2874-8. PMC: 51342. DOI: 10.1073/pnas.88.7.2874. View

10.

Wickner S, Hoskins J, McKenney K . Monomerization of RepA dimers by heat shock proteins activates binding to DNA replication origin. Proc Natl Acad Sci U S A. 1991; 88(18):7903-7. PMC: 52413. DOI: 10.1073/pnas.88.18.7903. View

11.

Wishart D, Sykes B, Richards F . The chemical shift index: a fast and simple method for the assignment of protein secondary structure through NMR spectroscopy. Biochemistry. 1992; 31(6):1647-51. DOI: 10.1021/bi00121a010. View

12.

Langer T, Lu C, Echols H, Flanagan J, Hayer M, Hartl F . Successive action of DnaK, DnaJ and GroEL along the pathway of chaperone-mediated protein folding. Nature. 1992; 356(6371):683-9. DOI: 10.1038/356683a0. View

13.

Bork P, Sander C, Valencia A, Bukau B . A module of the DnaJ heat shock proteins found in malaria parasites. Trends Biochem Sci. 1992; 17(4):129. DOI: 10.1016/0968-0004(92)90319-5. View

14.

Clore G, Bax A, Gronenborn A . Stereospecific assignment of beta-methylene protons in larger proteins using 3D 15N-separated Hartmann-Hahn and 13C-separated rotating frame Overhauser spectroscopy. J Biomol NMR. 1991; 1(1):13-22. DOI: 10.1007/BF01874566. View

15.

Piotto M, Saudek V, SKLENAR V . Gradient-tailored excitation for single-quantum NMR spectroscopy of aqueous solutions. J Biomol NMR. 1992; 2(6):661-5. DOI: 10.1007/BF02192855. View

16.

Grzesiek S, Vuister G, Bax A . A simple and sensitive experiment for measurement of JCC couplings between backbone carbonyl and methyl carbons in isotopically enriched proteins. J Biomol NMR. 1993; 3(4):487-93. DOI: 10.1007/BF00176014. View

17.

Bai Y, Milne J, Mayne L, Englander S . Primary structure effects on peptide group hydrogen exchange. Proteins. 1993; 17(1):75-86. PMC: 3438223. DOI: 10.1002/prot.340170110. View

18.

Wall D, Zylicz M, Georgopoulos C . The conserved G/F motif of the DnaJ chaperone is necessary for the activation of the substrate binding properties of the DnaK chaperone. J Biol Chem. 1995; 270(5):2139-44. DOI: 10.1074/jbc.270.5.2139. View

19.

Hill R, Flanagan J, Prestegard J . 1H and 15N magnetic resonance assignments, secondary structure, and tertiary fold of Escherichia coli DnaJ(1-78). Biochemistry. 1995; 34(16):5587-96. DOI: 10.1021/bi00016a033. View

20.

Wishart D, Bigam C, Yao J, Abildgaard F, Dyson H, Oldfield E . 1H, 13C and 15N chemical shift referencing in biomolecular NMR. J Biomol NMR. 1995; 6(2):135-40. DOI: 10.1007/BF00211777. View