3.4.25.2 A188S clpY mutant, the mutant shows altered interaction with SulA substrates, wild-type and mutant, and altered induction by arabinose or glutamate compared to the wild-type, overview 712278 3.4.25.2 DELTA111-239 2 Gly linker, amidolytic ativity is 60-80% of the activity of the wild-type enzyme, caseinolytic activity is 60-80% of the activity of the wild-type enzyme, activity with SulA-MBP fusion protein is less than 20% of the activity of the wild-type enzyme, ATPase activity is 60-80% of the activity of the wild-type enzyme 647883 3.4.25.2 DELTA137-150 2 Gly linker, amidolytic ativity, caseinolytic activity, activity with SulA-MBP fusion protein and ATPase activity are unchanged 647883 3.4.25.2 DELTA175-209 2 Gly linker, amidolytic ativity, caseinolytic activity, and ATPase activity are unchanged, activity with the SalU-MBP fusion protein is less than 20% of the activity of the wild-type enzyme 647883 3.4.25.2 DELTA423-443 5 Gly insertion, no amidolytic activity, no activity with casein and SulA-MBP fusion protein, no ATPase activity 647883 3.4.25.2 DELTA83-92 hydrolysis of casein, SulA-MBP or benzyloxycarbonyl-GGL-7-amido-4-methylcoumarin is less than 20% of the activity of the wild-type enzyme 647890 3.4.25.2 DELTA86-91 hydrolysis of casein, SulA-MBP or benzyloxycarbonyl-GGL-7-amido-4-methylcoumarin is unchanged 647890 3.4.25.2 DELTA88-92 3 Gly linker, amidolytic ativity, caseinolytic activity, activity with SulA-MBP fusion protein and ATPase activity are less than 20% of the activity of the wild-type enzyme 647883 3.4.25.2 DELTA89-92 1 Gly linker, amidolytic ativity is 40-60% of the activity of the wild-type enzyme, caseinolytic activity is 40-60% of the activity of the wild-type enzyme, activity with SulA-MBP fusion protein is less than 20% of the activity of the wild-type enzyme, ATPase activity is 40-60% of the activity of the wild-type enzyme 647883 3.4.25.2 E193L/E194L clpY mutant, the mutant shows altered interaction with SulA substrates, wild-type and mutant, and altered induction by arabinose or glutamate compared to the wild-type, overview 712278 3.4.25.2 E266Q amidolytic ativity, caseinolytic activity, activity with SulA-MBP fusion protein and ATPase activity are unchanged 647883 3.4.25.2 E266Q/E385 amidolytic ativity, caseinolytic activity, activity with SulA-MBP fusion protein and ATPase activity are unchanged 647883 3.4.25.2 E286Q amidolytic ativity is 40-60% of the activity of the wild-type enzyme, caseinolytic activity is 40-60% of the activity of the wild-type enzyme, activity with SulA-MBP fusion protein is 40-60% of the activity of the wild-type enzyme, ATPase activity is unchanged 647883 3.4.25.2 E321Q amidolytic ativity, caseinolytic activity, activity with SulA-MBP fusion protein and ATPase activity is less than 20% of the activity of the wild-type enzyme 647883 3.4.25.2 E325E amidolytic ativity, caseinolytic activity, activity with SulA-MBP fusion protein and ATPase activity are less than 20% of the activity of the wild-type enzyme. Crystal structure of the mutant complex is nearly identical to then active complex 647883 3.4.25.2 E436K/D437K amidolytic ativity is 60-80% of the activity of the wild-type enzyme, caseinolytic activity is unchanged, activity with SulA-MBP fusion protein is less than 20% of the activity of the wild-type enzyme, ATPase activity is unchanged 647883 3.4.25.2 E61C clpQ mutant 712278 3.4.25.2 E88Q amidolytic ativity is 20-40% of the activity of the wild-type enzyme, caseinolytic activity is less than 20% of the activity of the wild-type enzyme, activity with SulA-MBP fusion protein is less than 20% of the activity of the wild-type enzyme, ATPase activity is unchanged 647883 3.4.25.2 E88Q/E266Q amidolytic ativity is 20-40% of the activity of the wild-type enzyme, caseinolytic activity is less than 20% of the activity of the wild-type enzyme, activity with SulA-MBP fusion protein is less than 20% of the activity of the wild-type enzyme, ATPase activity is unchanged 647883 3.4.25.2 E95W amidolytic activity, activity with casein and ATPase activity are unchanged, activity with SulA-MBP fusion protein is 20-40% of the activity of the wild-type enzyme 647883 3.4.25.2 G90P mutation of the GYVG motif residues affects protein unfolding, ATP hydrolysis, affinity for ADP, and interaction of HslU and HslV, overview, the mutant shows 41% reduced ATP hydrolysis activity compared to wild-type HslU 665631 3.4.25.2 G93A amidolytic ativity is 20-40% of the activity of the wild-type enzyme, caseinolytic activity is 20-40% of the activity of the wild-type enzyme, activity with SulA-MBP fusion protein is less than 20% of the activity of the wild-type enzyme, ATPase activity is 40-60% of the activity of the wild-type enzyme 647883 3.4.25.2 G93A mutation of the GYVG motif residues affects protein unfolding, ATP hydrolysis, affinity for ADP, and interaction of HslU and HslV, overview 665631 3.4.25.2 G93P mutation of the GYVG motif residues affects protein unfolding, ATP hydrolysis, affinity for ADP, and interaction of HslU and HslV, overview 665631 3.4.25.2 I186N clpY mutant, the mutant does not interact with SulA compared to the wild-type ClpY 712278 3.4.25.2 I312W amidolytic ativity, caseinolytic activity, activity with SulA-MBP fusion protein and ATPase activity are higher than the wild-type activities 647883 3.4.25.2 Ins(435,436) 5 Gly insertion, no amidolytic activity, no activity with casein and SulA-MBP fusion protein, no ATPase activity 647883 3.4.25.2 K28A hydrolysis of casein, SulA-MBP or benzyloxycarbonyl-GGL-7-amido-4-methylcoumarin is less than 20% of the activity of the wild-type enzyme 647890 3.4.25.2 K80T amidolytic ativity is 20-40% of the activity of the wild-type enzyme, caseinolytic activity is 40-60% of the activity of the wild-type enzyme, activity with SulA-MBP fusion protein is unchanged, ATPase activity is unchanged 647883 3.4.25.2 L199Q clpY mutant, the mutant shows altered interaction with SulA substrates, wild-type and mutant, and altered induction by arabinose or glutamate compared to the wild-type, overview. SulA accumulates in the bacterial cells that express ClpY 712278 3.4.25.2 L199Q substitution in I-domain of subunit HslU. Mutation does not alter the structure of the AAA+ ring or its interactions with HslV but increases I-domain susceptibility to limited endoproteolysis. The mutation increases the rate of ATP-hydrolysis substantially, results in slower degradation of some proteins but faster degradation of other substrates, and markedly changes the preference of HslUV for initiating degradation at the N-terminus or C-terminus of model substrates 755561 3.4.25.2 L88A the mutation leads to a tighter binding between HslV and HslU and a dramatic stimulation of both the proteolytic and ATPase activities. Furthermore, the HslV mutant shows a more than 7fold increase of basal hydrolytic activities toward small peptides and unstructured proteins 734475 3.4.25.2 L88F the muattion increases the peptidolytic activity of HslV in both the absence and presence of HslU and stimulates the ATPase activity of HslU more than wild type HslV 734475 3.4.25.2 L88G the HslV mutant shows a marked increase of basal hydrolytic activities toward small peptides and unstructured proteins 734475 3.4.25.2 L88S the HslV mutant shows a marked increase of basal hydrolytic activities toward small peptides and unstructured proteins 734475 3.4.25.2 L88W the muattion increases the peptidolytic activity of HslV in both the absence and presence of HslU and stimulates the ATPase activity of HslU more than wild type HslV 734475 3.4.25.2 M187I clpY mutant, the mutant shows altered interaction with SulA substrates, wild-type and mutant, and altered induction by arabinose or glutamate compared to the wild-type, overview 712278 3.4.25.2 additional information clpQ+Y+ promoter is fused to a lacZ reporter gene. The transcriptional or translational clpQ+::lacZ fusion gene is each crossed into lambda phage. The lambdaclpQ+::lacZ+, a transcriptional fusion gene, is used to form lysogens in the wild-type, rpoH or/and rpoS mutants. Upon shifting the temperature up from 30°C to 42°C, the wild-type transcriptional lambdaclpQ+::lacZ+ demonstrates an increased beta-galactosidase activity, overview. RpoH itself regulates clpQ+Y+ gene expression. The clpQ+Y+ message carries a conserved 71 bp at the 5'-untranslated region that is predicted to form the stem-loop structure by analysis of its RNA secondary structure 713162 3.4.25.2 additional information construction of deletion mutants DELTA175-209linker, DELTA175-209GG, and DELTA108-243GG 720996 3.4.25.2 additional information construction of mixed dodecamers having varied numbers of HslV and T1A subunits, and of a series of HslV dodecamers containing different numbers of active sites showing that HslV with only 6 active sites is sufficient to support full catalytic activity, a further reduction of the number of active sites leads to a proportional decrease in activity. Substrate-mediated stabilization of the HslV-HslU interaction remains unchanged until the number of the active sites is decreased to 6 but is gradually compromised upon further reduction. Deletion of Thr1 causes a dramatic increase in affinity between HslV and HslU 712397 3.4.25.2 additional information construction of truncation mutants lacking the substrate binding residues 137-209 of ClpY 712278 3.4.25.2 N141L/N142L the ClpY loop 1 mutant is defective in complete degradation of SulA 712278 3.4.25.2 N205K clpY mutant, the mutant shows altered interaction with SulA substrates, wild-type and mutant, and altered induction by arabinose or glutamate compared to the wild-type, overview 712278 3.4.25.2 Q148L/Q149L/Q150L the ClpY loop 1 mutant shows altered substrate recognition and binding, but shows normal activity similar to that of the wild-type ClpY 712278 3.4.25.2 Q198L/Q200L clpY mutant, the mutant shows altered interaction with SulA substrates, wild-type and mutant, and altered induction by arabinose or glutamate compared to the wild-type, overview 712278 3.4.25.2 Q311_I312insGGGGG 5 Gly insertion, amidolytic ativity, caseinolytic activity and activity with SulA-MBP fusion protein are less than 20% of the activity of the wild-type enzyme, ATPase activity is 20-40% of the activity of the wild-type enzyme 647883 3.4.25.2 R35A hydrolysis of casein, SulA-MBP or benzyloxycarbonyl-GGL-7-amido-4-methylcoumarin is less than 20% of the activity of the wild-type enzyme 647890 3.4.25.2 R393A amidolytic ativity, caseinolytic activity, activity with SulA-MBP fusion protein and ATPase activity is less than 20% of the activity of the wild-type enzyme 647883 3.4.25.2 R86A the mutant shows little peptidolytic activity compared to the wild type 734475 3.4.25.2 R86D hydrolysis of casein, SulA-MBP or benzyloxycarbonyl-GGL-7-amido-4-methylcoumarin is less than 20% of the activity of the wild-type enzyme 647890 3.4.25.2 R86G ATP inhibits the degradation of unfolded proteins by HslV. This inhibitory effect of ATP is markedly diminished by substitution of the Arg86 residue located in the apical pore of HslV with Gly 688941 3.4.25.2 R89A the mutant shows little peptidolytic activity compared to the wild type 734475 3.4.25.2 R89A/K90A hydrolysis of casein, SulA-MBP or benzyloxycarbonyl-GGL-7-amido-4-methylcoumarin is less than 20% of the activity of the wild-type enzyme 647890 3.4.25.2 R89D hydrolysis of casein, SulA-MBP or benzyloxycarbonyl-GGL-7-amido-4-methylcoumarin is 40-60% of the activity of the wild-type enzyme 647890 3.4.25.2 R89D/K90E hydrolysis of casein and SulA-MBP is less than 20% of the activity of the wild-type enzyme, hydrolysis of benzyloxycarbonyl-GGL-7-amido-4-methylcoumarin is higher than that of the wild-type enzyme 647890 3.4.25.2 S103A 50% of the activity of the wild-type enzyme with benzyloxycarbonyl-GGL-7-amido-4-methylcoumarin in presence of the ATPase component HslU 647871 3.4.25.2 S124A 3% of the activity of the wild-type enzyme with benzyloxycarbonyl-GGL-7-amido-4-methylcoumarin in presence of the ATPase component HslU 647871 3.4.25.2 S143A 95% of the activity of the wild-type enzyme with benzyloxycarbonyl-GGL-7-amido-4-methylcoumarin in presence of the ATPase component HslU 647871 3.4.25.2 S172A 1% of the activity of the wild-type enzyme with benzyloxycarbonyl-GGL-7-amido-4-methylcoumarin in presence of the ATPase component HslU 647871 3.4.25.2 S5A 124% of the activity of the wild-type enzyme with benzyloxycarbonyl-GGL-7-amido-4-methylcoumarin in presence of the ATPase component HslU 647871 3.4.25.2 T387_E388insGGGGG 5 Gly insertion, amidolytic ativity is unchanged, caseinolytic activity is 60-80% of the activity of the wild-type enzyme, ATPase activity is unchanged 647883 3.4.25.2 V112A hydrolysis of casein, SulA-MBP or benzyloxycarbonyl-GGL-7-amido-4-methylcoumarin is less than 20% of the activity of the wild-type enzyme 647890 3.4.25.2 V92A mutation of the GYVG motif residues affects protein unfolding, ATP hydrolysis, affinity for ADP, and interaction of HslU and HslV, overview 665631 3.4.25.2 V92G amidolytic activity, activity with casein and ATPase activity are unchanged, activity with SulA-MBP fusion protein is less than 20% of the activity of the wild-type enzyme 647883 3.4.25.2 V92I mutation of the GYVG motif residues affects protein unfolding, ATP hydrolysis, affinity for ADP, and interaction of HslU and HslV, overview 665631 3.4.25.2 V92S mutation of the GYVG motif residues affects protein unfolding, ATP hydrolysis, affinity for ADP, and interaction of HslU and HslV, overview 665631 3.4.25.2 Y91A mutation of the GYVG motif residues affects protein unfolding, ATP hydrolysis, affinity for ADP, and interaction of HslU and HslV, overview 665631 3.4.25.2 Y91A site-directed mutagenesis of the HslU GYVG pore loop, the mutant shows no remaining activity, the mutant hydrolyzes ATP and stimulates HslV peptidase activity 720996 3.4.25.2 Y91F mutation of the GYVG motif residues affects protein unfolding, ATP hydrolysis, affinity for ADP, and interaction of HslU and HslV, overview 665631 3.4.25.2 Y91F site-directed mutagenesis of the HslU GYVG pore loop, the mutant shows reduced activity, the mutant hydrolyzes ATP and stimulates HslV peptidase activity 720996 3.4.25.2 Y91G amidolytic ativity is 40-60% of the activity of the wild-type enzyme, caseinolytic activity is 40-60% of the activity of the wild-type enzyme, activity with SulA-MBP fusion protein is less than 20% of the activity of the wild-type enzyme, ATPase activity is unchanged 647883 3.4.25.2 Y91S mutation of the GYVG motif residues affects protein unfolding, ATP hydrolysis, affinity for ADP, and interaction of HslU and HslV, overview 665631