Role of DnaJ G/F-rich Domain in Conformational Recognition and Binding of Protein Substrates

Overview

Journal J Biol Chem

Specialty Biochemistry

Date 2010 Aug 24

PMID 20729526

Citations 38

Authors

Judit Perales-Calvo

Arturo Muga

Fernando Moro

Affiliations

Soon will be listed here.

Abstract

DnaJ from Escherichia coli is a Type I Hsp40 that functions as a cochaperone of DnaK (Hsp70), stimulating its ATPase activity and delivering protein substrates. How DnaJ binds protein substrates is still poorly understood. Here we have studied the role of DnaJ G/F-rich domain in binding of several substrates with different conformational properties (folded, partially (un)folded and unfolded). Using partial proteolysis we find that RepE, a folded substrate, contacts a wide DnaJ area that involves part of the G/F-rich region and Zn-binding domain. Deletion of G/F-rich region hampers binding of native RepE and reduced the affinity for partially (un)folded substrates. However, binding of completely unfolded substrates is independent on the G/F-rich region. These data indicate that DnaJ distinguishes the substrate conformation and is able to adapt the use of the G/F-rich region to form stable substrate complexes.

Citing Articles

Force-regulated chaperone activity of BiP/ERdj3 is opposite to their homologs DnaK/DnaJ.

Banerjee S, Chowdhury D, Chakraborty S, Haldar S Protein Sci. 2024; 33(7):e5068.

PMID: 38864739 PMC: 11168073. DOI: 10.1002/pro.5068.

Tandem repeats of highly bioluminescent NanoLuc are refolded noncanonically by the Hsp70 machinery.

Apostolidou D, Zhang P, Pandya D, Bock K, Liu Q, Yang W Protein Sci. 2024; 33(2):e4895.

PMID: 38284490 PMC: 10804678. DOI: 10.1002/pro.4895.

Mining the Genome for Virulence Genes: A Functional-Based Approach to Discover Novel Loci Mediating Blue Mold Decay of Apple Fruit.

Luciano-Rosario D, Peng H, Gaskins V, Fonseca J, Keller N, Jurick 2nd W J Fungi (Basel). 2023; 9(11).

PMID: 37998873 PMC: 10672711. DOI: 10.3390/jof9111066.

Real-time single-molecule observation of chaperone-assisted protein folding.

Marzano N, Paudel B, van Oijen A, Ecroyd H Sci Adv. 2022; 8(50):eadd0922.

PMID: 36516244 PMC: 9750156. DOI: 10.1126/sciadv.add0922.

Disease-associated mutations within the yeast DNAJB6 homolog Sis1 slow conformer-specific substrate processing and can be corrected by the modulation of nucleotide exchange factors.

Bhadra A, Rau M, Daw J, Fitzpatrick J, Weihl C, True H Nat Commun. 2022; 13(1):4570.

PMID: 35931773 PMC: 9355953. DOI: 10.1038/s41467-022-32318-9.

References

Mokranjac D, Bourenkov G, Hell K, Neupert W, Groll M . Structure and function of Tim14 and Tim16, the J and J-like components of the mitochondrial protein import motor. EMBO J. 2006; 25(19):4675-85. PMC: 1590002. DOI: 10.1038/sj.emboj.7601334. View

Cajo G, Horne B, Kelley W, Schwager F, Georgopoulos C, Genevaux P . The role of the DIF motif of the DnaJ (Hsp40) co-chaperone in the regulation of the DnaK (Hsp70) chaperone cycle. J Biol Chem. 2006; 281(18):12436-44. DOI: 10.1074/jbc.M511192200. View

Feifel B, Schonfeld H, Christen P . D-peptide ligands for the co-chaperone DnaJ. J Biol Chem. 1998; 273(20):11999-2002. DOI: 10.1074/jbc.273.20.11999. View

Zylicz M, Yamamoto T, McKittrick N, Sell S, Georgopoulos C . Purification and properties of the dnaJ replication protein of Escherichia coli. J Biol Chem. 1985; 260(12):7591-8. View

Rudiger S, Schneider-Mergener J, Bukau B . Its substrate specificity characterizes the DnaJ co-chaperone as a scanning factor for the DnaK chaperone. EMBO J. 2001; 20(5):1042-50. PMC: 145471. DOI: 10.1093/emboj/20.5.1042. View

Karzai A, McMacken R . A bipartite signaling mechanism involved in DnaJ-mediated activation of the Escherichia coli DnaK protein. J Biol Chem. 1996; 271(19):11236-46. DOI: 10.1074/jbc.271.19.11236. View

Laufen T, Mayer M, Beisel C, Klostermeier D, Mogk A, Reinstein J . Mechanism of regulation of hsp70 chaperones by DnaJ cochaperones. Proc Natl Acad Sci U S A. 1999; 96(10):5452-7. PMC: 21880. DOI: 10.1073/pnas.96.10.5452. View

Mehl A, Heskett L, Neal K . A GrpE mutant containing the NH(2)-terminal "tail" region is able to displace bound polypeptide substrate from DnaK. Biochem Biophys Res Commun. 2001; 282(2):562-9. DOI: 10.1006/bbrc.2001.4567. View

Gruschus J, Han C, Greener T, Ferretti J, Greene L, Eisenberg E . Structure of the functional fragment of auxilin required for catalytic uncoating of clathrin-coated vesicles. Biochemistry. 2004; 43(11):3111-9. DOI: 10.1021/bi0354740. View

10.

Schlieker C, Tews I, Bukau B, Mogk A . Solubilization of aggregated proteins by ClpB/DnaK relies on the continuous extraction of unfolded polypeptides. FEBS Lett. 2004; 578(3):351-6. DOI: 10.1016/j.febslet.2004.11.051. View

11.

Li J, Qian X, Sha B . The crystal structure of the yeast Hsp40 Ydj1 complexed with its peptide substrate. Structure. 2003; 11(12):1475-83. DOI: 10.1016/j.str.2003.10.012. View

12.

Montgomery D, Morimoto R, Gierasch L . Mutations in the substrate binding domain of the Escherichia coli 70 kDa molecular chaperone, DnaK, which alter substrate affinity or interdomain coupling. J Mol Biol. 1999; 286(3):915-32. DOI: 10.1006/jmbi.1998.2514. View

13.

Moro F, Fernandez V, Muga A . Interdomain interaction through helices A and B of DnaK peptide binding domain. FEBS Lett. 2002; 533(1-3):119-23. DOI: 10.1016/s0014-5793(02)03752-3. View

14.

Lee S, Fan C, Younger J, Ren H, Cyr D . Identification of essential residues in the type II Hsp40 Sis1 that function in polypeptide binding. J Biol Chem. 2002; 277(24):21675-82. DOI: 10.1074/jbc.M111075200. View

15.

Sadler I, Chiang A, Kurihara T, Rothblatt J, Way J, Silver P . A yeast gene important for protein assembly into the endoplasmic reticulum and the nucleus has homology to DnaJ, an Escherichia coli heat shock protein. J Cell Biol. 1989; 109(6 Pt 1):2665-75. PMC: 2115965. DOI: 10.1083/jcb.109.6.2665. View

16.

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

17.

Cheetham M, Caplan A . Structure, function and evolution of DnaJ: conservation and adaptation of chaperone function. Cell Stress Chaperones. 1998; 3(1):28-36. PMC: 312945. DOI: 10.1379/1466-1268(1998)003<0028:sfaeod>2.3.co;2. View

18.

Rist W, Jorgensen T, Roepstorff P, Bukau B, Mayer M . Mapping temperature-induced conformational changes in the Escherichia coli heat shock transcription factor sigma 32 by amide hydrogen exchange. J Biol Chem. 2003; 278(51):51415-21. DOI: 10.1074/jbc.M307160200. View

19.

Wall D, Zylicz M, Georgopoulos C . The NH2-terminal 108 amino acids of the Escherichia coli DnaJ protein stimulate the ATPase activity of DnaK and are sufficient for lambda replication. J Biol Chem. 1994; 269(7):5446-51. View

20.

Wawrzynow A, Zylicz M . Divergent effects of ATP on the binding of the DnaK and DnaJ chaperones to each other, or to their various native and denatured protein substrates. J Biol Chem. 1995; 270(33):19300-6. DOI: 10.1074/jbc.270.33.19300. View