P0A6Y8 (DNAK_ECOLI) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
Chaperone protein DnaK

Alternative name(s):
HSP70
Heat shock 70 kDa protein
Heat shock protein 70

Gene names

Name:	dnaK
Synonyms:	groP, grpF, seg
Ordered Locus Names:	b0014, JW0013

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	638 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	Plays an essential role in the initiation of phage lambda DNA replication, where it acts in an ATP-dependent fashion with the DnaJ protein to release lambda O and P proteins from the preprimosomal complex. DnaK is also involved in chromosomal DNA replication, possibly through an analogous interaction with the DnaA protein. Also participates actively in the response to hyperosmotic shock. HAMAP-Rule MF_00332
Subcellular location	Cytoplasm. Cell inner membrane; Peripheral membrane protein Ref.16.
Post-translational modification	Autophosphorylated; GrpE inhibits the autophosphorylation. Ref.11 Ref.12 Ref.13
Disruption phenotype	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 all at 43 degrees) and strain background. Ref.15
Sequence similarities	Belongs to the heat shock protein 70 family.

Ontologies

Keywords
Biological process	DNA replication Stress response
Cellular component	Cell inner membrane Cell membrane Cytoplasm Membrane
Ligand	ATP-binding Nucleotide-binding
Molecular function	Chaperone
PTM	Acetylation Phosphoprotein
Technical term	3D-structure Complete proteome Direct protein sequencing Reference proteome
Gene Ontology (GO)
Biological_process	DNA replication Inferred from electronic annotation. Source: UniProtKB-KW cellular response to unfolded protein Inferred from direct assay PubMed 2203539PubMed 7900997. Source: EcoCyc chaperone cofactor-dependent protein refolding Inferred from direct assay PubMed 2203539PubMed 7900997. Source: EcoCyc chaperone mediated protein folding requiring cofactor Inferred from genetic interaction PubMed 10458167. Source: EcoCyc protein complex assembly Inferred from direct assay PubMed 2525129PubMed 2525130. Source: EcoCyc protein complex disassembly Inferred from direct assay PubMed 2525130PubMed 2543679. Source: EcoCyc protein unfolding Inferred from direct assay PubMed 20953191. Source: EcoCyc response to heat Inferred from direct assay PubMed 7023474PubMed 7900997. Source: EcoCyc
Cellular_component	cytoplasm Inferred from direct assay PubMed 16858726. Source: UniProtKB membrane Inferred from direct assay PubMed 16858726. Source: UniProtKB plasma membrane Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular_function	ADP binding Inferred from direct assay PubMed 19439666PubMed 7776367. Source: EcoCyc ATP binding Inferred from direct assay PubMed 7776367. Source: EcoCyc unfolded protein binding Inferred from direct assay PubMed 9145101. Source: EcoCyc zinc ion binding Inferred from direct assay PubMed 11985624. Source: EcoliWiki
Complete GO annotation...

Binary interactions

With	Entry	#Exp.	IntAct	Notes
aspA	P0AC38	2	EBI-542092,EBI-544200
dnaJ	P08622	5	EBI-542092,EBI-545285
fabF	P0AAI5	2	EBI-542092,EBI-542783
fbaA	P0AB71	2	EBI-542092,EBI-370916
gloA	P0AC81	2	EBI-542092,EBI-551143
rne	P21513	9	EBI-542092,EBI-549958
ydhR	P0ACX3	2	EBI-542092,EBI-544817
ygcP	Q46906	2	EBI-542092,EBI-557727

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Initiator methionine

1

1

Removed Ref.5 Ref.6 Ref.11

Chain

2 – 638

637

Chaperone protein DnaK HAMAP-Rule MF_00332

PRO_0000078458

Amino acid modifications

Modified residue

70

1

N6-succinyllysine Ref.18

Modified residue

109

1

N6-acetyllysine Ref.17

Modified residue

199

1

Phosphothreonine; by autocatalysis Ref.12 Ref.13

Modified residue

245

1

N6-acetyllysine; alternate Ref.17

Modified residue

245

1

N6-succinyllysine; alternate Ref.18

Modified residue

246

1

N6-succinyllysine Ref.18

Modified residue

304

1

N6-acetyllysine; alternate Ref.17

Modified residue

304

1

N6-succinyllysine; alternate Ref.18

Modified residue

359

1

N6-succinyllysine Ref.18

Modified residue

421

1

N6-acetyllysine Ref.17

Modified residue

502

1

N6-succinyllysine Ref.18

Modified residue

528

1

N6-succinyllysine Ref.18

Modified residue

556

1

N6-acetyllysine Ref.17

Modified residue

587

1

N6-succinyllysine Ref.18

Experimental info

Mutagenesis

32

1

G → D in SEG-1 and dnaK756(TS); confers temperature sensitivity. Ref.9 Ref.10

Mutagenesis

436

1

V → I in SEG-2; confers temperature sensitivity. Ref.10

Mutagenesis

455

1

G → D in dnaK756(TS); confers temperature sensitivity. Ref.9

Mutagenesis

468

1

G → D in dnaK756(TS); confers temperature sensitivity. Ref.9

Secondary structure

1

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638

Helix Strand Turn

Details...

Beta strand

4 – 8

5

Beta strand

11 – 20

10

Beta strand

23 – 26

4

Beta strand

34 – 37

4

Beta strand

40 – 42

3

Beta strand

48 – 51

4

Helix

52 – 56

5

Turn

57 – 60

4

Helix

62 – 64

3

Beta strand

65 – 67

3

Helix

69 – 71

3

Turn

72 – 74

3

Beta strand

76 – 79

4

Helix

80 – 86

7

Beta strand

90 – 95

6

Beta strand

99 – 105

7

Beta strand

108 – 110

3

Helix

112 – 131

20

Beta strand

137 – 142

6

Helix

148 – 160

13

Beta strand

164 – 170

7

Helix

171 – 180

10

Beta strand

186 – 195

10

Beta strand

200 – 212

13

Beta strand

214 – 224

11

Helix

229 – 247

19

Helix

252 – 254

3

Helix

256 – 272

17

Turn

273 – 275

3

Beta strand

277 – 288

12

Beta strand

290 – 301

12

Helix

302 – 315

14

Helix

318 – 326

9

Helix

331 – 333

3

Beta strand

335 – 341

7

Helix

342 – 345

4

Helix

347 – 357

11

Beta strand

363 – 365

3

Turn

367 – 369

3

Helix

370 – 383

14

Beta strand

384 – 386

3

Beta strand

388 – 392

5

Beta strand

394 – 397

4

Beta strand

399 – 403

5

Turn

404 – 406

3

Beta strand

407 – 412

6

Beta strand

414 – 416

3

Beta strand

417 – 430

14

Beta strand

435 – 444

10

Beta strand

445 – 447

3

Helix

448 – 450

3

Beta strand

451 – 460

10

Beta strand

467 – 469

3

Beta strand

472 – 478

7

Beta strand

480 – 482

3

Beta strand

484 – 490

7

Turn

491 – 493

3

Beta strand

496 – 501

6

Helix

509 – 521

13

Helix

523 – 553

31

Helix

554 – 556

3

Helix

559 – 577

19

Helix

581 – 594

14

Helix

596 – 601

6

Sequences

	Sequence		Length	Mass (Da)
	P0A6Y8 [UniParc]. Last modified January 23, 2007. Version 2. Checksum: 5A8589B21D7CD9C1	FASTA	638	69,115
	10 20 30 40 50 60 MGKIIGIDLG TTNSCVAIMD GTTPRVLENA EGDRTTPSII AYTQDGETLV GQPAKRQAVT 70 80 90 100 110 120 NPQNTLFAIK RLIGRRFQDE EVQRDVSIMP FKIIAADNGD AWVEVKGQKM APPQISAEVL 130 140 150 160 170 180 KKMKKTAEDY LGEPVTEAVI TVPAYFNDAQ RQATKDAGRI AGLEVKRIIN EPTAAALAYG 190 200 210 220 230 240 LDKGTGNRTI AVYDLGGGTF DISIIEIDEV DGEKTFEVLA TNGDTHLGGE DFDSRLINYL 250 260 270 280 290 300 VEEFKKDQGI DLRNDPLAMQ RLKEAAEKAK IELSSAQQTD VNLPYITADA TGPKHMNIKV 310 320 330 340 350 360 TRAKLESLVE DLVNRSIEPL KVALQDAGLS VSDIDDVILV GGQTRMPMVQ KKVAEFFGKE 370 380 390 400 410 420 PRKDVNPDEA VAIGAAVQGG VLTGDVKDVL LLDVTPLSLG IETMGGVMTT LIAKNTTIPT 430 440 450 460 470 480 KHSQVFSTAE DNQSAVTIHV LQGERKRAAD NKSLGQFNLD GINPAPRGMP QIEVTFDIDA 490 500 510 520 530 540 DGILHVSAKD KNSGKEQKIT IKASSGLNED EIQKMVRDAE ANAEADRKFE ELVQTRNQGD 550 560 570 580 590 600 HLLHSTRKQV EEAGDKLPAD DKTAIESALT ALETALKGED KAAIEAKMQE LAQVSQKLME 610 620 630 IAQQQHAQQQ TAGADASANN AKDDDVVDAE FEEVKDKK

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Major heat shock gene of Drosophila and the Escherichia coli heat-inducible dnaK gene are homologous." Bardwell J.C.A., Craig E.A. Proc. Natl. Acad. Sci. U.S.A. 81:848-852(1984) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
[2]	"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.
[3]	"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.
[4]	"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.
[5]	"Precursor of mitochondrial aspartate aminotransferase synthesized in Escherichia coli is complexed with heat-shock protein DnaK." Schmid D., Jaussi R., Christen P. Eur. J. Biochem. 208:699-704(1992) [PubMed] [Europe PMC] [Abstract] Cited for: PROTEIN SEQUENCE OF 2-15.
[6]	"Comparing the predicted and observed properties of proteins encoded in the genome of Escherichia coli K-12." Link A.J., Robison K., Church G.M. Electrophoresis 18:1259-1313(1997) [PubMed] [Europe PMC] [Abstract] Cited for: PROTEIN SEQUENCE OF 2-13. Strain: K12 / EMG2.
[7]	"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] OF 590-638.
[8]	"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] OF 628-638.
[9]	"DNA sequence analysis of the dnaK gene of Escherichia coli B and of two dnaK genes carrying the temperature-sensitive mutations dnaK7(Ts) and dnaK756(Ts)." Miyazaki T., Tanaka S., Fujita H., Itikawa H. J. Bacteriol. 174:3715-3722(1992) [PubMed] [Europe PMC] [Abstract] Cited for: MUTAGENESIS OF GLY-32; GLY-455 AND GLY-468. Strain: B.
[10]	"Involvement of DnaK protein in mini-F plasmid replication: temperature-sensitive seg mutations are located in the dnaK gene." Ezaki B., Ogura T., Mori H., Niki H., Hiraga S. Mol. Gen. Genet. 218:183-189(1989) [PubMed] [Europe PMC] [Abstract] Cited for: MUTAGENESIS OF GLY-32 AND VAL-436. Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
[11]	"Identification of phosphoproteins in Escherichia coli." Freestone P., Grant S., Toth I., Norris V. Mol. Microbiol. 15:573-580(1995) [PubMed] [Europe PMC] [Abstract] Cited for: PHOSPHORYLATION, PROTEIN SEQUENCE OF 2-11.
[12]	"DnaK as a thermometer: threonine-199 is site of autophosphorylation and is critical for ATPase activity." McCarty J.S., Walker G.C. Proc. Natl. Acad. Sci. U.S.A. 88:9513-9517(1991) [PubMed] [Europe PMC] [Abstract] Cited for: PHOSPHORYLATION AT THR-199.
[13]	"Inhibition of DnaK autophosphorylation by heat shock proteins and polypeptide substrates." Panagiotidis C.A., Burkholder W.F., Gaitanaris G.A., Gragerov A., Gottesman M.E., Silverstein S.J. J. Biol. Chem. 269:16643-16647(1994) [PubMed] [Europe PMC] [Abstract] Cited for: PHOSPHORYLATION AT THR-199.
[14]	"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.
[15]	"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.
[16]	"Protein complexes of the Escherichia coli cell envelope." Stenberg F., Chovanec P., Maslen S.L., Robinson C.V., Ilag L., von Heijne G., Daley D.O. J. Biol. Chem. 280:34409-34419(2005) [PubMed] [Europe PMC] [Abstract] Cited for: SUBCELLULAR LOCATION. Strain: BL21-DE3.
[17]	"Lysine acetylation is a highly abundant and evolutionarily conserved modification in Escherichia coli." Zhang J., Sprung R., Pei J., Tan X., Kim S., Zhu H., Liu C.F., Grishin N.V., Zhao Y. Mol. Cell. Proteomics 8:215-225(2009) [PubMed] [Europe PMC] [Abstract] Cited for: ACETYLATION [LARGE SCALE ANALYSIS] AT LYS-109; LYS-245; LYS-304; LYS-421 AND LYS-556, MASS SPECTROMETRY. Strain: K12 / JW1106 and K12 / MG1655 / ATCC 47076.
[18]	"Identification of lysine succinylation as a new post-translational modification." Zhang Z., Tan M., Xie Z., Dai L., Chen Y., Zhao Y. Nat. Chem. Biol. 7:58-63(2011) [PubMed] [Europe PMC] [Abstract] Cited for: SUCCINYLATION AT LYS-70; LYS-245; LYS-246; LYS-304; LYS-359; LYS-502; LYS-528 AND LYS-587. Strain: K12.
[19]	"Structural analysis of substrate binding by the molecular chaperone DnaK." Zhu X., Zhao X., Burkholder W.F., Gragerov A., Ogata C.M., Gottesman M.E., Hendrickson W.A. Science 272:1606-1614(1996) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS) OF 389-607.
[20]	"NMR solution structure of the 21 kDa chaperone protein DnaK substrate binding domain: a preview of chaperone-protein interaction." Wang H., Kurochkin A.V., Pang Y., Hu W., Flynn G.C., Zuiderweg E.R.P. Biochemistry 37:7929-7940(1998) [PubMed] [Europe PMC] [Abstract] Cited for: STRUCTURE BY NMR OF 387-562.
[21]	"Structural insights into substrate binding by the molecular chaperone DnaK." Pellecchia M., Montgomery D.L., Stevens S.Y., Vander Kooi C.W., Feng H.P., Gierasch L.M., Zuiderweg E.R.P. Nat. Struct. Biol. 7:298-303(2000) [PubMed] [Europe PMC] [Abstract] Cited for: STRUCTURE BY NMR OF 394-508.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

K01298 Genomic DNA. Translation: AAA23694.1.
U00096 Genomic DNA. Translation: AAC73125.1.
AP009048 Genomic DNA. Translation: BAB96589.1.
D10765 Genomic DNA. Translation: BAA01595.1.
M12565 Genomic DNA. Translation: AAA23692.1.

PIR

IQECDK. A03311.

RefSeq

NP_414555.1. NC_000913.2.
YP_488320.1. NC_007779.1.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1BPR	NMR	-	A	386-561	[»]
1DG4	NMR	-	A	393-507	[»]
1DKG	X-ray	2.80	D	1-383	[»]
1DKX	X-ray	2.00	A	389-607	[»]
1DKY	X-ray	2.80	A/B	389-607	[»]
1DKZ	X-ray	2.00	A	389-607	[»]
1Q5L	NMR	-	A	393-506	[»]
2BPR	NMR	-	A	386-561	[»]
2KHO	NMR	-	A	1-605	[»]
3DPO	X-ray	2.10	A/B	389-607	[»]
3DPP	X-ray	2.50	A/B	389-607	[»]
3DPQ	X-ray	2.60	A/B/E/F	389-607	[»]
3QNJ	X-ray	2.28	A/B	389-607	[»]
4B9Q	X-ray	2.40	A/B/C/D	1-605	[»]
4E81	X-ray	1.90	A/B	389-607	[»]

ProteinModelPortal

P0A6Y8.

SMR

P0A6Y8. Positions 4-606.

ModBase

Search...

Protein-protein interaction databases

DIP

DIP-35751N.

IntAct

P0A6Y8. 365 interactions.

MINT

MINT-7259066.

STRING

511145.b0014.

Protein family/group databases

TCDB

1.A.33.1.2. cation channel-forming heat shock protein-70 (Hsp70) family.

PTM databases

PhosSite

P010484.

2D gel databases

SWISS-2DPAGE

P0A6Y8.

Proteomic databases

PaxDb

P0A6Y8.

PRIDE

P0A6Y8.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Genome annotation databases

EnsemblBacteria

AAC73125; AAC73125; b0014.
BAB96589; BAB96589; BAB96589.

GeneID

12930526.
944750.

KEGG

ecj:Y75_p0014.
eco:b0014.

PATRIC

32115121. VBIEscCol129921_0012.

Organism-specific databases

EchoBASE

EB0237.

EcoGene

EG10241. dnaK.

Phylogenomic databases

eggNOG

COG0443.

HOGENOM

HOG000228136.

KO

K04043.

OMA

PYKNINP.

ProtClustDB

PRK00290.

Enzyme and pathway databases

BioCyc

EcoCyc:EG10241-MONOMER.
ECOL316407:JW0013-MONOMER.

Gene expression databases

Genevestigator

P0A6Y8.

Family and domain databases

HAMAP

MF_00332. DnaK.

InterPro

IPR012725. Chaperone_DnaK.
IPR018181. Heat_shock_70_CS.
IPR013126. Hsp_70_fam.
[Graphical view]

Pfam

PF00012. HSP70. 1 hit.
[Graphical view]

PRINTS

PR00301. HEATSHOCK70.

TIGRFAMs

TIGR02350. prok_dnaK. 1 hit.

PROSITE

PS00297. HSP70_1. 1 hit.
PS00329. HSP70_2. 1 hit.
PS01036. HSP70_3. 1 hit.
[Graphical view]

ProtoNet

Search...

Other

EvolutionaryTrace

P0A6Y8.

Entry information

Entry name

DNAK_ECOLI

Accession

Primary (citable) accession number: P0A6Y8
Secondary accession number(s): P04475

Entry history

Integrated into UniProtKB/Swiss-Prot:	August 13, 1987
Last sequence update:	January 23, 2007
Last modified:	May 1, 2013
This is version 94 of the entry and version 2 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program