P0A6H5 (HSLU_ECOLI) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
ATP-dependent protease ATPase subunit HslU

Alternative name(s):
Heat shock protein HslU
Unfoldase HslU

Gene names

Name:	hslU
Synonyms:	htpI
Ordered Locus Names:	b3931, JW3902

Organism

Escherichia coli (strain K12)

Taxonomic identifier

83333 [NCBI]

Taxonomic lineage

Bacteria › Proteobacteria › Gammaproteobacteria › Enterobacteriales › Enterobacteriaceae › Escherichia

Protein attributes

Sequence length	443 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	ATPase subunit of a proteasome-like degradation complex; this subunit has chaperone activity. The binding of ATP and its subsequent hydrolysis by HslU are essential for unfolding of protein substrates subsequently hydrolyzed by HslV. HslU recognizes the N-terminal part of its protein substrates and unfolds these before they are guided to HslV for hydrolysis. Ref.6 Ref.7 Ref.9 Ref.10 Ref.12 Ref.13 Ref.14
Subunit structure	A double ring-shaped homohexamer of HslV is capped on each side by a ring-shaped HslU homohexamer. The assembly of the HslU/HslV complex is dependent on binding of ATP.
Subcellular location	Cytoplasm HAMAP MF_00249.
Induction	By heat shock. Ref.1
Sequence similarities	Belongs to the ClpX chaperone family. HslU subfamily.
Biophysicochemical properties	Kinetic parameters: Arc is a repressor protein, Arc-MYL-st11 is a hyperstable variant of Arc. KM=0.31 mM for ATP (in the absence of HslV) Ref.6 Ref.14 KM=0.28 mM for ATP (in the presence of HslV) KM=5.2 µM for Arc-MYL-st11 (at 37 degrees Celsius) Vmax=57 pmol/min/mg enzyme (in the absence of HslV) Vmax=213 pmol/min/mg enzyme (in the presence of HslV) Temperature dependence: Optimum temperature is 55 degrees Celsius.

Ontologies

Keywords
Biological process	Stress response
Cellular component	Cytoplasm
Ligand	ATP-binding Nucleotide-binding
Molecular function	Chaperone
Technical term	3D-structure Complete proteome Reference proteome
Gene Ontology (GO)
Biological process	response to heat Inferred from expression pattern. Source: EcoliWiki
Cellular component	HslUV protease complex Inferred from electronic annotation. Source: InterPro membrane Inferred from direct assay. Source: UniProtKB
Molecular function	ATP binding Inferred from electronic annotation. Source: UniProtKB-KW ATPase activity Inferred from electronic annotation. Source: InterPro identical protein binding Inferred from physical interaction. Source: IntAct peptidase activity, acting on L-amino acid peptides Inferred from electronic annotation. Source: InterPro
Complete GO annotation...

Binary interactions

With	Entry	#Exp.	IntAct	Notes
itself		2	EBI-369317,EBI-369317
hslV	P0A7B8	7	EBI-369317,EBI-552265

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Chain

1 – 443

443

ATP-dependent protease ATPase subunit HslU HAMAP MF_00249

PRO_0000160500

Regions

Nucleotide binding

60 – 65

6

ATP HAMAP MF_00249

Sites

Binding site

18

1

ATP; via amide nitrogen and carbonyl oxygen

Binding site

256

1

ATP

Binding site

321

1

ATP

Binding site

393

1

ATP

Experimental info

Mutagenesis

63

1

K → T: Can neither bind nor hydrolyze ATP. Do not form multimers, but stays as monomer. Ref.5

Mutagenesis

80

1

K → T: Some effect on protease activity. Ref.17

Mutagenesis

88

1

E → Q: Severly reduced protease activity. Ref.17

Mutagenesis

91

1

Y → G: Partial loss of protease activity. Ref.17

Mutagenesis

92

1

V → G: Partial loss of protease activity. Ref.17

Mutagenesis

93

1

G → A: Almost no protease or ATP hydrolysis activity. Ref.17

Mutagenesis

95

1

E → W: Partial loss of protease activity. Ref.17

Mutagenesis

262

1

C → V: No effect on ATP hydrolysis. Can support HslV-mediated proteolysis at wild-type levels. Ref.11

Mutagenesis

266

1

E → Q: No effect. Ref.17

Mutagenesis

286

1

E → Q: Reduced protease activity. Ref.17

Mutagenesis

288

1

C → V: No ATP hydrolysis activity. Binds ATP with lower affinity than wild-type. Can support HslV-mediated proteolysis to some extent. Ref.11

Mutagenesis

312

1

I → W: No effect. Ref.17

Mutagenesis

321

1

E → Q: Complete loss of activity. Ref.17

Mutagenesis

325

1

R → E: Complete loss of activity. Forms wild-type complexes with HslV and is able to bind ATP. Ref.17

Mutagenesis

385

1

E → K: No effect. Ref.17

Mutagenesis

393

1

R → A: Complete loss of activity. Ref.17

Mutagenesis

436

1

E → K: Partial loss of protease activity; when associated with K-437.

Mutagenesis

437

1

D → K: Partial loss of protease activity; when associated with K-436.

Secondary structure

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443

Helix Strand Turn

Details...

Helix

6 – 13

8

Turn

14 – 16

3

Helix

21 – 37

17

Helix

42 – 45

4

Beta strand

53 – 56

4

Helix

63 – 74

12

Beta strand

78 – 82

5

Helix

83 – 86

4

Beta strand

89 – 91

3

Helix

98 – 118

21

Helix

120 – 129

10

Helix

132 – 135

4

Beta strand

148 – 152

5

Helix

153 – 164

12

Beta strand

171 – 173

3

Beta strand

214 – 216

3

Helix

218 – 233

16

Helix

236 – 250

15

Beta strand

252 – 256

5

Turn

257 – 261

5

Helix

269 – 286

18

Beta strand

289 – 292

4

Beta strand

295 – 298

4

Helix

299 – 301

3

Beta strand

303 – 308

6

Beta strand

311 – 313

3

Helix

315 – 317

3

Helix

320 – 324

5

Beta strand

328 – 331

4

Helix

337 – 345

9

Helix

351 – 360

10

Turn

361 – 363

3

Beta strand

365 – 368

4

Helix

370 – 386

17

Helix

393 – 409

17

Helix

410 – 413

4

Beta strand

417 – 420

4

Helix

422 – 434

13

Helix

436 – 439

4

Sequences

	Sequence		Length	Mass (Da)
	P0A6H5 [UniParc]. Last modified March 29, 2005. Version 1. Checksum: D2188639EA9AEF4C	FASTA	443	49,594
	10 20 30 40 50 60 MSEMTPREIV SELDKHIIGQ DNAKRSVAIA LRNRWRRMQL NEELRHEVTP KNILMIGPTG 70 80 90 100 110 120 VGKTEIARRL AKLANAPFIK VEATKFTEVG YVGKEVDSII RDLTDAAVKM VRVQAIEKNR 130 140 150 160 170 180 YRAEELAEER ILDVLIPPAK NNWGQTEQQQ EPSAARQAFR KKLREGQLDD KEIEIDLAAA 190 200 210 220 230 240 PMGVEIMAPP GMEEMTSQLQ SMFQNLGGQK QKARKLKIKD AMKLLIEEEA AKLVNPEELK 250 260 270 280 290 300 QDAIDAVEQH GIVFIDEIDK ICKRGESSGP DVSREGVQRD LLPLVEGCTV STKHGMVKTD 310 320 330 340 350 360 HILFIASGAF QIAKPSDLIP ELQGRLPIRV ELQALTTSDF ERILTEPNAS ITVQYKALMA 370 380 390 400 410 420 TEGVNIEFTD SGIKRIAEAA WQVNESTENI GARRLHTVLE RLMEEISYDA SDLSGQNITI 430 440 DADYVSKHLD ALVADEDLSR FIL

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Sequence analysis of four new heat-shock genes constituting the hslTS/ibpAB and hslVU operons in Escherichia coli." Chuang S.E., Burland V., Plunkett G. III, Daniels D.L., Blattner F.R. Gene 134:1-6(1993) [PubMed: 8244018] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], INDUCTION. Strain: K12 / MG1655 / ATCC 47076.
[2]	"Analysis of the Escherichia coli genome. III. DNA sequence of the region from 87.2 to 89.2 minutes." Plunkett G. III, Burland V., Daniels D.L., Blattner F.R. Nucleic Acids Res. 21:3391-3398(1993) [PubMed: 8346018] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / MG1655 / ATCC 47076.
[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: 9278503] [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: 16738553] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
[5]	"Mutational analysis of the ATP-binding site in HslU, the ATPase component of HslVU protease in Escherichia coli." Shin D.H., Yoo S.J., Shim Y.K., Seol J.H., Kang M.S., Chung C.H. FEBS Lett. 398:151-154(1996) [PubMed: 8977096] [Abstract] Cited for: MUTAGENESIS OF LYS-63.
[6]	"Purification and characterization of the heat shock proteins HslV and HslU that form a new ATP-dependent protease in Escherichia coli." Yoo S.J., Seol J.H., Shin D.H., Rohrwild M., Kang M.-S., Tanaka K., Goldberg A.L., Chung C.H. J. Biol. Chem. 271:14035-14040(1996) [PubMed: 8662828] [Abstract] Cited for: FUNCTION, SUBSTRATE SPECIFICITY, BIOPHYSICOCHEMICAL PROPERTIES.
[7]	"HslV-HslU: A novel ATP-dependent protease complex in Escherichia coli related to the eukaryotic proteasome." Rohrwild M., Coux O., Huang H.-C., Moerschell R.P., Yoo S.J., Seol J.H., Chung C.H., Goldberg A.L. Proc. Natl. Acad. Sci. U.S.A. 93:5808-5813(1996) [PubMed: 8650174] [Abstract] Cited for: FUNCTION, CATALYTIC ACTIVITY.
[8]	"ATP binding, but not its hydrolysis, is required for assembly and proteolytic activity of the HslVU protease in Escherichia coli." Yoo S.J., Seol J.H., Seong I.S., Kang M.-S., Chung C.H. Biochem. Biophys. Res. Commun. 238:581-585(1997) [PubMed: 9299555] [Abstract] Cited for: EFFECTS OF ATP BINDING ON COMPLEX FORMATION.
[9]	"The heat-shock protein HslVU from Escherichia coli is a protein-activated ATPase as well as an ATP-dependent proteinase." Seol J.H., Yoo S.J., Shin D.H., Shim Y.K., Kang M.-S., Goldberg A.L., Chung C.H. Eur. J. Biochem. 247:1143-1150(1997) [PubMed: 9288941] [Abstract] Cited for: FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY.
[10]	"Synergistic roles of HslVU and other ATP-dependent proteases in controlling in vivo turnover of sigma32 and abnormal proteins in Escherichia coli." Kanemori M., Nishihara K., Yanagi H., Yura T. J. Bacteriol. 179:7219-7225(1997) [PubMed: 9393683] [Abstract] Cited for: FUNCTION, CATALYTIC ACTIVITY.
[11]	"Effects of the cys mutations on structure and function of the ATP-dependent HslVU protease in Escherichia coli. The Cys287 to Val mutation in HslU uncouples the ATP-dependent proteolysis by HslvU from ATP hydrolysis." Yoo S.J., Kim H.H., Shin D.H., Lee C.S., Seong I.S., Seol J.H., Shimbara N., Tanaka K., Chung C.H. J. Biol. Chem. 273:22929-22935(1998) [PubMed: 9722513] [Abstract] Cited for: MUTAGENESIS OF CYS-262 AND CYS-288.
[12]	"ATP-dependent degradation of SulA, a cell division inhibitor, by the HslVU protease in Escherichia coli." Seong I.S., Oh J.Y., Yoo S.J., Seol J.H., Chung C.H. FEBS Lett. 456:211-214(1999) [PubMed: 10452560] [Abstract] Cited for: FUNCTION, CATALYTIC ACTIVITY.
[13]	"Marked instability of the sigma(32) heat shock transcription factor at high temperature. Implications for heat shock regulation." Kanemori M., Yanagi H., Yura T. J. Biol. Chem. 274:22002-22007(1999) [PubMed: 10419524] [Abstract] Cited for: FUNCTION, CATALYTIC ACTIVITY, CHARACTERIZATION.
[14]	"Nucleotide-dependent substrate recognition by the AAA+ HslUV protease." Burton R.E., Baker T.A., Sauer R.T. Nat. Struct. Mol. Biol. 12:245-251(2005) [PubMed: 15696175] [Abstract] Cited for: FUNCTION, BIOPHYSICOCHEMICAL PROPERTIES.
[15]	"HslVU ATP-dependent protease utilizes maximally six among twelve threonine active sites during proteolysis." Lee J.W., Park E., Jeong M.S., Jeon Y.J., Eom S.H., Seol J.H., Chung C.H. J. Biol. Chem. 284:33475-33484(2009) [PubMed: 19801685] [Abstract] Cited for: REACTION MECHANISM.
[16]	"The structures of HslU and the ATP-dependent protease HslU-HslV." Bochtler M., Hartmann C., Song H.K., Bourenkov G.P., Bartunik H.D., Huber R. Nature 403:800-805(2000) [PubMed: 10693812] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.81 ANGSTROMS) OF COMPLEXES WITH ATP AND HSLV.
[17]	"Mutational studies on HslU and its docking mode with HslV." Song H.K., Hartmann C., Ramachandran R., Bochtler M., Behrendt R., Moroder L., Huber R. Proc. Natl. Acad. Sci. U.S.A. 97:14103-14108(2000) [PubMed: 11114186] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.8 ANGSTROMS) OF 2-176 OF APOENZYME AND IN COMPLEXES WITH HSLV AND ATP, MUTAGENESIS OF LYS-80; GLU-88; TYR-91; VAL-92; GLY-93; GLU-95; GLU-266; GLU-286; ILE-312; GLU-321; ARG-325; GLU-385 AND ARG-393.
[18]	"Nucleotide-dependent conformational changes in a protease-associated ATPase HslU." Wang J., Song J.J., Seong I.S., Franklin M.C., Kamtekar S., Eom S.H., Chung C.H. Structure 9:1107-1116(2001) [PubMed: 11709174] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.8 ANGSTROMS) OF 2-443 IN COMPLEX WITH ADP.
[19]	"Correction of X-ray intensities from an HslV-HslU co-crystal containing lattice-translocation defects." Wang J., Rho S.H., Park H.H., Eom S.H. Acta Crystallogr. D 61:932-941(2005) [PubMed: 15983416] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (4.16 ANGSTROMS) IN COMPLEX WITH B.SUBTILIS HSLV AND ADP.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

L19201 Genomic DNA. Translation: AAB03063.1.
U00096 Genomic DNA. Translation: AAC76913.1.
AP009048 Genomic DNA. Translation: BAE77379.1.

PIR

JT0761.

RefSeq

NP_418366.1. NC_000913.2.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1DO0	X-ray	3.00	A/B/C/D/E/F	2-443	[»]
1DO2	X-ray	4.00	A/B/C/D	2-443	[»]
1E94	X-ray	2.80	E/F	2-443	[»]
1G4A	X-ray	3.00	E/F	1-443	[»]
1G4B	X-ray	7.00	E/F/K/L	1-443	[»]
1HQY	X-ray	2.80	E/F	2-443	[»]
1HT1	X-ray	2.80	E/F/G/I	2-443	[»]
1HT2	X-ray	2.80	E/F/G/H	2-443	[»]
1YYF	X-ray	4.16	A/B	1-443	[»]

ProteinModelPortal

P0A6H5.

SMR

P0A6H5. Positions 1-443.

ModBase

Search...

Protein-protein interaction databases

DIP

DIP-31855N.

IntAct

P0A6H5. 48 interactions.

2D gel databases

SWISS-2DPAGE

P0A6H5.

ECO2DBASE

D048.5. 6TH EDITION.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Genome annotation databases

EnsemblBacteria

EBESCT00000003987; EBESCP00000003987; EBESCG00000003262.
EBESCT00000018172; EBESCP00000017463; EBESCG00000017227.

GeneID

948430.

GenomeReviews

Gene locus JW3902 in contig AP009048_GR.
Gene locus b3931 in contig U00096_GR.

KEGG

ecj:JW3902.
eco:b3931.

PATRIC

32123377. VBIEscCol129921_4049.

Organism-specific databases

EchoBASE

EB1827.

EcoGene

EG11881. hslU.

Phylogenomic databases

eggNOG

COG1220.

GeneTree

EBGT00050000010205.

HOGENOM

HBG745965.

OMA

EASKFTE.

PhylomeDB

P0A6H5.

ProtClustDB

PRK05201.

Enzyme and pathway databases

BioCyc

EcoCyc:EG11881-MONOMER.

Gene expression databases

Genevestigator

P0A6H5.

Family and domain databases

HAMAP

MF_00249. HslU.
[Tree]

InterPro

IPR003593. ATPase_AAA+_core.
IPR013093. ATPase_AAA-2.
IPR003959. ATPase_AAA_core.
IPR019489. Clp_ATPase_C.
IPR004491. HslU.
[Graphical view]

KO

K03667.

PANTHER

PTHR11262:SF3. HslU. 1 hit.

Pfam

PF00004. AAA. 1 hit.
PF07724. AAA_2. 1 hit.
PF10431. ClpB_D2-small. 1 hit.
[Graphical view]

SMART

SM00382. AAA. 1 hit.
SM01086. ClpB_D2-small. 1 hit.
[Graphical view]

TIGRFAMs

TIGR00390. HslU. 1 hit.

ProtoNet

Search...

Entry information

Entry name

HSLU_ECOLI

Accession

Primary (citable) accession number: P0A6H5
Secondary accession number(s): P32168, Q2M8M7

Entry history

Integrated into UniProtKB/Swiss-Prot:	March 29, 2005
Last sequence update:	March 29, 2005
Last modified:	January 25, 2012
This is version 71 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program