P0A7B8 (HSLV_ECOLI) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
ATP-dependent protease subunit HslV
EC=3.4.25.2

Alternative name(s):
Heat shock protein HslV

Gene names

Name:	hslV
Synonyms:	htpO, yiiC
Ordered Locus Names:	b3932, JW3903

Organism

Escherichia coli (strain K12)

Taxonomic identifier

83333 [NCBI]

Taxonomic lineage

Bacteria › Proteobacteria › Gammaproteobacteria › Enterobacteriales › Enterobacteriaceae › Escherichia

Protein attributes

Sequence length	176 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	Protease subunit of a proteasome-like degradation complex believed to be a general protein degrading machinery. The complex has been shown to be involved in the specific degradation of heat shock induced transcription factors such as RpoH and SulA. In addition, small hydrophobic peptides are also hydrolyzed by HslV. HslV has weak protease activity even in the absence of HslU, but this activity is induced more than 100-fold in the presence of HslU. HslU recognizes protein substrates and unfolds these before guiding them to HslV for hydrolysis. HslV is not believed to degrade folded proteins. Ref.7 Ref.8 Ref.10 Ref.11 Ref.14 Ref.15 Ref.16
Catalytic activity	ATP-dependent cleavage of peptide bonds with broad specificity. Ref.8 Ref.10 Ref.11 Ref.14 Ref.15
Enzyme regulation	Allosterically activated by HslU binding Probable. HAMAP MF_00248
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. Ref.18
Subcellular location	Cytoplasm HAMAP MF_00248.
Induction	By heat shock. Ref.1
Sequence similarities	Belongs to the peptidase T1B family. HslV subfamily.
Caution	Ref.5 sequence is supposed to originate from rat but, based on sequence similarity, it seems that this is a case of bacterial contamination from E.coli.
Biophysicochemical properties	Kinetic parameters: Arc is a repressor protein, Arc-MYL-st11 is a hyperstable variant of Arc. KM=5.2 µM for Arc-MYL-st11 (at 37 degrees Celsius) Ref.16 Temperature dependence: Optimum temperature is 55 degrees Celsius.

Ontologies

Keywords
Biological process	Stress response
Cellular component	Cytoplasm
Ligand	Metal-binding Sodium
Molecular function	Hydrolase Protease Threonine protease
Technical term	3D-structure Allosteric enzyme Complete proteome Direct protein sequencing Reference proteome
Gene Ontology (GO)
Biological process	proteolysis involved in cellular protein catabolic process Inferred from electronic annotation. Source: InterPro response to heat Inferred from expression pattern. Source: EcoliWiki
Cellular component	HslUV protease complex Inferred from electronic annotation. Source: InterPro cytosol Inferred from direct assay. Source: UniProtKB proteasome core complex Inferred from electronic annotation. Source: InterPro
Molecular function	identical protein binding Inferred from physical interaction. Source: IntAct metal ion binding Inferred from electronic annotation. Source: UniProtKB-KW threonine-type endopeptidase activity Inferred from electronic annotation. Source: UniProtKB-KW
Complete GO annotation...

Binary interactions

With	Entry	#Exp.	IntAct	Notes
itself		2	EBI-552265,EBI-552265
hslU	P0A6H5	7	EBI-552265,EBI-369317

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Initiator methionine

1

1

Removed HAMAP MF_00248

Chain

2 – 176

175

ATP-dependent protease subunit HslV HAMAP MF_00248

PRO_0000148105

Sites

Active site

2

1

Ref.12

Metal binding

157

1

Sodium; via carbonyl oxygen

Metal binding

160

1

Sodium; via carbonyl oxygen

Metal binding

163

1

Sodium; via carbonyl oxygen

Experimental info

Mutagenesis

2

1

T → S: 80% reduced protease activity in the absence of HslU. Almost no effect in the presence of HslU. Ref.12

Mutagenesis

2

1

T → V: No protease activity. Ref.12

Mutagenesis

3

1

T → S or V: 80% reduced protease activity. Ref.12

Mutagenesis

6

1

S → A: No effect. Ref.12

Mutagenesis

104

1

S → A: 50% reduced protease activity. Ref.12

Mutagenesis

125

1

S → A: Almost no protease activity. Ref.12

Mutagenesis

144

1

S → A: No effect. Ref.12

Mutagenesis

160

1

C → A or S: No protease activity. Cannot form complexes with HslU. Ref.13

Mutagenesis

173

1

S → A: Almost no protease activity. Ref.12

Secondary structure

1

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

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.

.

.

.

176

Helix Strand Turn

Details...

Beta strand

4 – 7

4

Beta strand

12 – 17

6

Beta strand

23 – 25

3

Turn

39 – 42

4

Turn

51 – 54

4

Helix

55 – 68

14

Helix

74 – 77

4

Helix

80 – 83

4

Beta strand

86 – 88

3

Beta strand

106 – 108

3

Turn

109 – 111

3

Beta strand

112 – 114

3

Turn

117 – 119

3

Helix

127 – 138

12

Helix

145 – 159

15

Beta strand

168 – 173

6

Sequences

	Sequence		Length	Mass (Da)
	P0A7B8 [UniParc]. Last modified January 23, 2007. Version 2. Checksum: 3B35E01F51486965	FASTA	176	19,093
	10 20 30 40 50 60 MTTIVSVRRN GHVVIAGDGQ ATLGNTVMKG NVKKVRRLYN DKVIAGFAGG TADAFTLFEL 70 80 90 100 110 120 FERKLEMHQG HLVKAAVELA KDWRTDRMLR KLEALLAVAD ETASLIITGN GDVVQPENDL 130 140 150 160 170 IAIGSGGPYA QAAARALLEN TELSAREIAE KALDIAGDIC IYTNHFHTIE ELSYKA

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]	"Molecular cloning of a novel gene involved in serotonin receptor-mediated signal transduction in rat stomach." Ohya S., Takii T., Yamazaki H., Matsumori M., Onozaki K., Watanabe M., Imaizumi Y. FEBS Lett. 401:252-258(1997) [PubMed: 9013898] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-125.
[6]	Dai K., Xu Y., Lutkenhaus J. Submitted (APR-1993) to the EMBL/GenBank/DDBJ databases Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-88.
[7]	"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.
[8]	"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, PROTEIN SEQUENCE OF N-TERMINUS.
[9]	"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.
[10]	"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.
[11]	"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.
[12]	"Mutagenesis of two N-terminal Thr and five Ser residues in HslV, the proteolytic component of the ATP-dependent HslVU protease." Yoo S.J., Shim Y.K., Seong I.S., Seol J.H., Kang M.-S., Chung C.H. FEBS Lett. 412:57-60(1997) [PubMed: 9257689] [Abstract] Cited for: MUTAGENESIS OF THR-2; THR-3; SER-6; SER-104; SER-125; SER-144 AND SER-173, ACTIVE SITE.
[13]	"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-160.
[14]	"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.
[15]	"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.
[16]	"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.
[17]	"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.
[18]	"Crystal structure of heat shock locus V (HslV) from Escherichia coli." Bochtler M., Ditzel L., Groll M., Huber R. Proc. Natl. Acad. Sci. U.S.A. 94:6070-6074(1997) [PubMed: 9177170] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (3.8 ANGSTROMS), SUBUNIT.
[19]	"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 HSLU AND ATP ANALOG.
[20]	"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.
[21]	"Crystal structures of the HslVU peptidase-ATPase complex reveal an ATP-dependent proteolysis mechanism." Wang J., Song J.J., Franklin M.C., Kamtekar S., Im Y.J., Rho S.H., Seong I.S., Lee C.S., Chung C.H., Eom S.H. Structure 9:177-184(2001) [PubMed: 11250202] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (3.0 ANGSTROMS) OF 2-175.
[22]	"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 2-176.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

L19201 Genomic DNA. Translation: AAB03064.1.
U00096 Genomic DNA. Translation: AAC76914.1.
AP009048 Genomic DNA. Translation: BAE77378.1.
D89965 mRNA. Translation: BAA14040.1. Sequence problems.
L14281 Genomic DNA. No translation available.

PIR

JT0760.

RefSeq

NP_418367.1. NC_000913.2.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1E94	X-ray	2.80	A/B/C/D	2-176	[»]
1G4A	X-ray	3.00	A/B/C/D	2-175	[»]
1G4B	X-ray	7.00	M/N/O/P	2-175	[»]
1HQY	X-ray	2.80	A/B/C/D	2-176	[»]
1HT1	X-ray	2.80	A/B/C/D/V/X/Y/Z	2-176	[»]
1HT2	X-ray	2.80	A/B/C/D/I/J/K/L	2-176	[»]
1NED	X-ray	3.80	A/B/C	2-176	[»]

ProteinModelPortal

P0A7B8.

SMR

P0A7B8. Positions 2-175.

ModBase

Search...

Protein-protein interaction databases

DIP

DIP-35866N.

IntAct

P0A7B8. 34 interactions.

MINT

MINT-1218940.

Protein family/group databases

MEROPS

T01.006.

2D gel databases

SWISS-2DPAGE

P0A7B8.

ECO2DBASE

G021.0. 6TH EDITION.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Genome annotation databases

EnsemblBacteria

EBESCT00000003716; EBESCP00000003716; EBESCG00000003037.
EBESCT00000017074; EBESCP00000016365; EBESCG00000016133.

GeneID

948429.

GenomeReviews

Gene locus JW3903 in contig AP009048_GR.
Gene locus b3932 in contig U00096_GR.

KEGG

ecj:JW3903.
eco:b3932.

PATRIC

32123379. VBIEscCol129921_4050.

Organism-specific databases

EchoBASE

EB1627.

EcoGene

EG11676. hslV.

Phylogenomic databases

eggNOG

COG5405.

GeneTree

EBGT00050000011263.

HOGENOM

HBG288822.

OMA

WRTDKYL.

PhylomeDB

P0A7B8.

ProtClustDB

PRK05456.

Enzyme and pathway databases

BioCyc

EcoCyc:EG11676-MONOMER.

Gene expression databases

Genevestigator

P0A7B8.

Family and domain databases

HAMAP

MF_00248. HslV.
[Tree]

InterPro

IPR022281. ATP-dep_Prtase_HsIV_su.
IPR001353. Proteasome_sua/b.
[Graphical view]

KO

K01419.

PANTHER

PTHR11599:SF38. PTHR11599:SF38. 1 hit.

Pfam

PF00227. Proteasome. 1 hit.
[Graphical view]

TIGRFAMs

TIGR03692. ATP_dep_HslV. 1 hit.

ProtoNet

Search...

Entry information

Entry name

HSLV_ECOLI

Accession

Primary (citable) accession number: P0A7B8
Secondary accession number(s): P31059, P97542, Q2M8M8

Entry history

Integrated into UniProtKB/Swiss-Prot:	June 7, 2005
Last sequence update:	January 23, 2007
Last modified:	January 25, 2012
This is version 70 of the entry and version 2 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program