P08622 (DNAJ_ECOLI) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
Chaperone protein DnaJ

Alternative name(s):
HSP40
Heat shock protein J

Gene names

Name:	dnaJ
Synonyms:	groP
Ordered Locus Names:	b0015, JW0014

Organism

Escherichia coli (strain K12) [Reference proteome] [HAMAP]

Taxonomic identifier

83333 [NCBI]

Taxonomic lineage

cellular organisms › Bacteria › Proteobacteria › Gammaproteobacteria › Enterobacteriales › Enterobacteriaceae › Escherichia › Escherichia coli

Protein attributes

Sequence length	376 AA.
Sequence status	Complete.
Sequence processing	The displayed sequence is further processed into a mature form.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	Interacts with DnaK and GrpE to disassemble a protein complex at the origins of replication of phage lambda and several plasmids. Participates actively in the response to hyperosmotic and heat shock by preventing the aggregation of stress-denatured proteins and by disaggregating proteins, also in an autonomous, DnaK-independent fashion. Unfolded proteins bind initially to DnaJ; upon interaction with the DnaJ-bound protein, DnaK hydrolyzes its bound ATP, resulting in the formation of a stable complex. GrpE releases ADP from DnaK; ATP binding to DnaK triggers the release of the substrate protein, thus completing the reaction cycle. Several rounds of ATP-dependent interactions between DnaJ, DnaK and GrpE are required for fully efficient folding. Ref.6 Ref.8 Ref.9 Ref.12
Cofactor	Binds 2 zinc ions per monomer.
Subunit structure	Homodimer.
Subcellular location	Cytoplasm HAMAP-Rule MF_01152.
Induction	By heat shock under the control of the HtpR regulatory protein. HAMAP-Rule MF_01152
Domain	The J domain is necessary and sufficient to stimulate DnaK ATPase activity. Zinc center 1 plays an important role in the autonomous, DnaK-independent chaperone activity of DnaJ. Zinc center 2 is essential for interaction with DnaK and for DnaJ activity. HAMAP-Rule MF_01152
Disruption phenotype	Single dnaJ and double dnaK-dnaJ disruption are non-essential; synthetic lethality is seen in a triple tig-dnaK-dnaJ disruption, although this depends on temperature (triple disruptions grow slowly at 20 and 34 degrees Celsius but not at 43 degrees) and strain background. Ref.10
Sequence similarities	Belongs to the DnaJ family. Contains 1 CR-type zinc finger. Contains 1 J domain.

Ontologies

Keywords
Biological process	DNA replication Stress response
Cellular component	Cytoplasm
Domain	Repeat Zinc-finger
Ligand	Metal-binding Zinc
Molecular function	Chaperone
Technical term	3D-structure Complete proteome Direct protein sequencing Reference proteome
Gene Ontology (GO)
Biological_process	DNA replication Inferred from mutant phenotype PubMed 2144273. Source: EcoliWiki protein refolding Inferred from direct assay PubMed 7559385. Source: EcoliWiki protein unfolding Inferred from direct assay PubMed 20953191. Source: EcoCyc response to heat Inferred from expression pattern PubMed 8349564. Source: EcoliWiki
Cellular_component	cytoplasm Inferred from direct assay PubMed 6220698. Source: EcoliWiki membrane Inferred from direct assay PubMed 3889001PubMed 6220698. Source: EcoliWiki
Molecular_function	ATP binding Inferred from electronic annotation. Source: InterPro protein disulfide isomerase activity Inferred from direct assay PubMed 7559385. Source: EcoliWiki unfolded protein binding Inferred from direct assay PubMed 7559385. Source: EcoliWiki zinc ion binding Inferred from direct assay PubMed 11985624PubMed 8662861. Source: EcoliWiki
Complete GO annotation...

Binary interactions

With	Entry	#Exp.	IntAct	Notes
dnaK	P0A6Y8	5	EBI-545285,EBI-542092

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Initiator methionine

1

1

Removed Ref.1

Chain

2 – 376

375

Chaperone protein DnaJ HAMAP-Rule MF_01152

PRO_0000070777

Regions

Domain

3 – 72

70

J

Repeat

144 – 151

8

CXXCXGXG motif HAMAP-Rule MF_01152

Repeat

161 – 168

8

CXXCXGXG motif HAMAP-Rule MF_01152

Repeat

183 – 190

8

CXXCXGXG motif HAMAP-Rule MF_01152

Repeat

197 – 204

8

CXXCXGXG motif HAMAP-Rule MF_01152

Zinc finger

131 – 209

79

CR-type HAMAP-Rule MF_01152

Compositional bias

77 – 114

38

Gly-rich HAMAP-Rule MF_01152

Sites

Metal binding

144

1

Zinc 1

Metal binding

147

1

Zinc 1

Metal binding

161

1

Zinc 2

Metal binding

164

1

Zinc 2

Metal binding

183

1

Zinc 2

Metal binding

186

1

Zinc 2

Metal binding

197

1

Zinc 1

Metal binding

200

1

Zinc 1

Experimental info

Mutagenesis

19 – 20

2

RE → AA: No effect. Ref.15

Mutagenesis

25

1

Y → A: Loss of activity.

Mutagenesis

26

1

K → A: Loss of activity.

Mutagenesis

27

1

R → A: No effect.

Mutagenesis

28

1

L → A: No effect.

Mutagenesis

29

1

A → G: No effect.

Mutagenesis

30 – 31

2

MK → AA: No effect.

Mutagenesis

32

1

Y → A: No effect.

Mutagenesis

33

1

H → Q: Loss of ability to stimulate DnaK ATPase activity. Ref.13

Mutagenesis

34

1

P → F: Loss of function.

Mutagenesis

35

1

D → N: Loss of ability to bind DnaK. Ref.14

Mutagenesis

36

1

R → A: Decrease in chaperone function.

Mutagenesis

37

1

N → A: Decrease in chaperone function.

Mutagenesis

38

1

Q → A: No effect.

Mutagenesis

41 – 42

2

KE → AA: No effect. Ref.15

Mutagenesis

44

1

E → A: No effect. Ref.15

Mutagenesis

46

1

K → A: No effect.

Mutagenesis

47

1

F → A: Loss of function.

Mutagenesis

48 – 49

2

KE → AA: No effect.

Mutagenesis

51 – 52

2

KE → AA: No effect. Ref.15

Mutagenesis

54

1

Y → A: No effect. Ref.15

Mutagenesis

55

1

E → A: No effect. Ref.15

Mutagenesis

58 – 59

2

TD → AA: No effect.

Mutagenesis

60 – 61

2

SQ → AA: No effect.

Mutagenesis

62 – 63

2

KR → AA: No effect.

Mutagenesis

67 – 68

2

DQ → AA: No effect. Ref.15

Mutagenesis

144

1

C → S: Loss of DnaK-independent chaperone activity; when associated with S-147; S-197 and S-200. Ref.16

Mutagenesis

147

1

C → S: Loss of DnaK-independent chaperone activity; when associated with S-144; S-197 and S-200. Ref.16

Mutagenesis

161

1

C → H: No effect on chaperone function; when associated with H-183. Ref.16 Ref.17

Mutagenesis

161

1

C → S: Loss of function; when associated with S-164; S-183 and S-186. Ref.16 Ref.17

Mutagenesis

164

1

C → H: No effect on chaperone function; when associated with H-183. Ref.16 Ref.17

Mutagenesis

164

1

C → S: Loss of function; when associated with S-161; S-183 and S-186. Ref.16 Ref.17

Mutagenesis

183

1

C → H: No effect on chaperone function. Same effect; when associated with H-161 or H-164. Ref.16 Ref.17

Mutagenesis

183

1

C → S: Loss of function; when associated with S-161; S-164 and S-186. Ref.16 Ref.17

Mutagenesis

186

1

C → H: No effect on chaperone function. Ref.16 Ref.17

Mutagenesis

186

1

C → S: Loss of function; when associated with S-161; S-164 and S-184. Ref.16 Ref.17

Mutagenesis

197

1

C → S: Loss of DnaK-independent chaperone activity; when associated with S-144; S-147 and S-200. Ref.16

Mutagenesis

200

1

C → S: Loss of DnaK-independent chaperone activity; when associated with S-144; S-147 and S-197. Ref.16

Secondary structure

1

.

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376

Helix Strand Turn

Details...

Turn

7 – 10

4

Beta strand

13 – 15

3

Helix

18 – 29

12

Turn

30 – 32

3

Turn

34 – 36

3

Turn

38 – 40

3

Helix

43 – 50

8

Turn

51 – 55

5

Helix

61 – 65

5

Turn

66 – 68

3

Turn

70 – 72

3

Turn

132 – 134

3

Beta strand

141 – 143

3

Helix

145 – 147

3

Turn

148 – 150

3

Beta strand

154 – 156

3

Turn

162 – 166

5

Beta strand

167 – 174

8

Beta strand

177 – 182

6

Turn

184 – 188

5

Beta strand

189 – 192

4

Beta strand

194 – 196

3

Helix

198 – 200

3

Beta strand

203 – 207

5

Sequences

	Sequence		Length	Mass (Da)
	P08622 [UniParc]. Last modified January 23, 2007. Version 3. Checksum: 05FA762EF9844532	FASTA	376	41,100
	10 20 30 40 50 60 MAKQDYYEIL GVSKTAEERE IRKAYKRLAM KYHPDRNQGD KEAEAKFKEI KEAYEVLTDS 70 80 90 100 110 120 QKRAAYDQYG HAAFEQGGMG GGGFGGGADF SDIFGDVFGD IFGGGRGRQR AARGADLRYN 130 140 150 160 170 180 MELTLEEAVR GVTKEIRIPT LEECDVCHGS GAKPGTQPQT CPTCHGSGQV QMRQGFFAVQ 190 200 210 220 230 240 QTCPHCQGRG TLIKDPCNKC HGHGRVERSK TLSVKIPAGV DTGDRIRLAG EGEAGEHGAP 250 260 270 280 290 300 AGDLYVQVQV KQHPIFEREG NNLYCEVPIN FAMAALGGEI EVPTLDGRVK LKVPGETQTG 310 320 330 340 350 360 KLFRMRGKGV KSVRGGAQGD LLCRVVVETP VGLNERQKQL LQELQESFGG PTGEHNSPRS 370 KSFFDGVKKF FDDLTR

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Nucleotide sequence of the Escherichia coli dnaJ gene and purification of the gene product." Ohki M., Tamura F., Nishimura S., Uchida H. J. Biol. Chem. 261:1778-1781(1986) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-6.
[2]	"The nucleotide sequence of the Escherichia coli K12 dnaJ+ gene. A gene that encodes a heat shock protein." Bardwell J.C.A., Tilly K., Craig E., King J., Zylicz M., Georgopoulos C. J. Biol. Chem. 261:1782-1785(1986) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA]. Strain: K12.
[3]	"Systematic sequencing of the Escherichia coli genome: analysis of the 0-2.4 min region." Yura T., Mori H., Nagai H., Nagata T., Ishihama A., Fujita N., Isono K., Mizobuchi K., Nakata A. Nucleic Acids Res. 20:3305-3308(1992) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12.
[4]	"The complete genome sequence of Escherichia coli K-12." Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V., Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F., Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B., Shao Y. Science 277:1453-1474(1997) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / MG1655 / ATCC 47076.
[5]	"Highly accurate genome sequences of Escherichia coli K-12 strains MG1655 and W3110." Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S., Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T. Mol. Syst. Biol. 2:E1-E5(2006) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
[6]	"Escherichia coli DnaJ and GrpE heat shock proteins jointly stimulate ATPase activity of DnaK." Liberek K., Marszalek J., Ang D., Georgopoulos C., Zylicz M. Proc. Natl. Acad. Sci. U.S.A. 88:2874-2878(1991) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION.
[7]	"Escherichia coli proteome analysis using the gene-protein database." VanBogelen R.A., Abshire K.Z., Moldover B., Olson E.R., Neidhardt F.C. Electrophoresis 18:1243-1251(1997) [PubMed] [Europe PMC] [Abstract] Cited for: IDENTIFICATION BY 2D-GEL.
[8]	"Successive and synergistic action of the Hsp70 and Hsp100 chaperones in protein disaggregation." Zietkiewicz S., Krzewska J., Liberek K. J. Biol. Chem. 279:44376-44383(2004) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION.
[9]	"Immediate response of the DnaK molecular chaperone system to heat shock." Siegenthaler R.K., Grimshaw J.P., Christen P. FEBS Lett. 562:105-110(2004) [PubMed] [Europe PMC] [Abstract] Cited for: ROLE IN HEAT-SHOCK RESPONSE.
[10]	"In vivo analysis of the overlapping functions of DnaK and trigger factor." Genevaux P., Keppel F., Schwager F., Langendijk-Genevaux P.S., Hartl F.U., Georgopoulos C. EMBO Rep. 5:195-200(2004) [PubMed] [Europe PMC] [Abstract] Cited for: DISRUPTION PHENOTYPE. Strain: K12 / MC4100 / ATCC 35695 / DSM 6574.
[11]	"Trigonal DnaK-DnaJ complex versus free DnaK and DnaJ: heat stress converts the former to the latter, and only the latter can do disaggregation in cooperation with ClpB." Watanabe Y.H., Yoshida M. J. Biol. Chem. 279:15723-15727(2004) [PubMed] [Europe PMC] [Abstract] Cited for: INTERACTION WITH DNAK.
[12]	"The DnaK-DnaJ-GrpE chaperone system activates inert wild type pi initiator protein of R6K into a form active in replication initiation." Zzaman S., Reddy J.M., Bastia D. J. Biol. Chem. 279:50886-50894(2004) [PubMed] [Europe PMC] [Abstract] Cited for: ROLE IN PLASMID DNA REPLICATION.
[13]	"The NH2-terminal 108 amino acids of the Escherichia coli DnaJ protein stimulate the ATPase activity of DnaK and are sufficient for lambda replication." Wall D., Zylicz M., Georgopoulos C. J. Biol. Chem. 269:5446-5451(1994) [PubMed] [Europe PMC] [Abstract] Cited for: MUTAGENESIS OF HIS-33.
[14]	"Interaction of the Hsp70 molecular chaperone, DnaK, with its cochaperone DnaJ." Suh W.-C., Burkholder W.F., Lu C.Z., Zhao X., Gottesman M.E., Gross C.A. Proc. Natl. Acad. Sci. U.S.A. 95:15223-15228(1998) [PubMed] [Europe PMC] [Abstract] Cited for: MUTAGENESIS OF ASP-35.
[15]	"Scanning mutagenesis identifies amino acid residues essential for the in vivo activity of the Escherichia coli DnaJ (Hsp40) J-domain." Genevaux P., Schwager F., Georgopoulos C., Kelley W.L. Genetics 162:1045-1053(2002) [PubMed] [Europe PMC] [Abstract] Cited for: MUTAGENESIS OF ARG-19; GLU-20; 25-TYR--GLN-38; LYS-41; GLU-42; GLU-44; 46-LYS--GLU-49; LYS-51; GLU-52; TYR-54; GLU-55; 58-THR--ARG-63; ASP-67 AND GLN-68.
[16]	"The roles of the two zinc binding sites in DnaJ." Linke K., Wolfram T., Bussemer J., Jakob U. J. Biol. Chem. 278:44457-44466(2003) [PubMed] [Europe PMC] [Abstract] Cited for: MUTAGENESIS OF CYS-144; CYS-147; CYS-161; CYS-164; CYS-183; CYS-186; CYS-197 AND CYS-200.
[17]	"Contributions of cysteine residues in Zn2 to zinc fingers and thiol-disulfide oxidoreductase activities of chaperone DnaJ." Shi Y.-Y., Tang W., Hao S.-F., Wang C.-C. Biochemistry 44:1683-1689(2005) [PubMed] [Europe PMC] [Abstract] Cited for: MUTAGENESIS OF CYS-161; CYS-164; CYS-183 AND CYS-186.
[18]	"NMR structure of the J-domain and the Gly/Phe-rich region of the Escherichia coli DnaJ chaperone." Pellechia M., Szyperski T., Wall D., Georgopoulos C., Wuethrich K. J. Mol. Biol. 260:236-250(1996) [PubMed] [Europe PMC] [Abstract] Cited for: STRUCTURE BY NMR OF 1-108.
[19]	"The influence of C-terminal extension on the structure of the 'J-domain' in E. coli DnaJ." Huang K., Flanagan J.M., Prestegard J.H. Protein Sci. 8:203-214(1999) [PubMed] [Europe PMC] [Abstract] Cited for: STRUCTURE BY NMR OF 1-105.
[20]	"Solution structure of the cysteine-rich domain of the Escherichia coli chaperone protein DnaJ." Martinez-Yamout M., Legge G.B., Zhang O., Wright P.E., Dyson H.J. J. Mol. Biol. 300:805-818(2000) [PubMed] [Europe PMC] [Abstract] Cited for: STRUCTURE BY NMR OF 131-209.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

M12544 Genomic DNA. Translation: AAA00009.1.
M12565 Genomic DNA. Translation: AAA23693.1.
U00096 Genomic DNA. Translation: AAC73126.1.
AP009048 Genomic DNA. Translation: BAB96590.1.

PIR

HHECDJ. A92572.

RefSeq

NP_414556.1. NC_000913.2.
YP_488321.1. NC_007779.1.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1BQ0	NMR	-	A	2-104	[»]
1BQZ	NMR	-	A	2-78	[»]
1EXK	NMR	-	A	131-209	[»]
1XBL	NMR	-	A	2-108	[»]

ProteinModelPortal

P08622.

SMR

P08622. Positions 2-78, 113-344.

ModBase

Search...

Protein-protein interaction databases

DIP

DIP-9460N.

IntAct

P08622. 94 interactions.

MINT

MINT-1220303.

STRING

511145.b0015.

Proteomic databases

PaxDb

P08622.

PRIDE

P08622.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Genome annotation databases

EnsemblBacteria

AAC73126; AAC73126; b0015.
BAB96590; BAB96590; BAB96590.

GeneID

12930731.
944753.

KEGG

ecj:Y75_p0015.
eco:b0015.

PATRIC

32115123. VBIEscCol129921_0013.

Organism-specific databases

EchoBASE

EB0236.

EcoGene

EG10240. dnaJ.

Phylogenomic databases

eggNOG

COG0484.

HOGENOM

HOG000226717.

KO

K03686.

OMA

HCHGRGK.

ProtClustDB

PRK10767.

Enzyme and pathway databases

BioCyc

EcoCyc:EG10240-MONOMER.
ECOL316407:JW0014-MONOMER.

Gene expression databases

Genevestigator

P08622.

Family and domain databases

Gene3D

1.10.287.110. 1 hit.
2.10.230.10. 1 hit.

HAMAP

MF_01152. DnaJ.

InterPro

IPR012724. DnaJ.
IPR002939. DnaJ_C.
IPR001623. DnaJ_domain.
IPR018253. DnaJ_domain_CS.
IPR008971. HSP40/DnaJ_pept-bd.
IPR001305. HSP_DnaJ_Cys-rich_dom.
[Graphical view]

Pfam

PF00226. DnaJ. 1 hit.
PF01556. DnaJ_C. 1 hit.
PF00684. DnaJ_CXXCXGXG. 1 hit.
[Graphical view]

PRINTS

PR00625. JDOMAIN.

SMART

SM00271. DnaJ. 1 hit.
[Graphical view]

SUPFAM

SSF46565. DnaJ_N. 1 hit.
SSF49493. HSP40_DnaJ_pep. 2 hits.
SSF57938. HSP_DnaJ_cys-rich. 1 hit.

TIGRFAMs

TIGR02349. DnaJ_bact. 1 hit.

PROSITE

PS00636. DNAJ_1. 1 hit.
PS50076. DNAJ_2. 1 hit.
PS51188. ZF_CR. 1 hit.
[Graphical view]

ProtoNet

Search...

Other

EvolutionaryTrace

P08622.

Entry information

Entry name

DNAJ_ECOLI

Accession

Primary (citable) accession number: P08622

Entry history

Integrated into UniProtKB/Swiss-Prot:	August 1, 1988
Last sequence update:	January 23, 2007
Last modified:	May 1, 2013
This is version 152 of the entry and version 3 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program