UniProt UniProt http://www.uniprot.org PspF-binding domain PspA1-144 and the PspA·F complex: New insights into the coiled-coil-dependent regulation of AAA+ proteins. PspF-binding domain PspA1-144 and the PspA·F complex: New insights into the coiled-coil-dependent regulation of AAA+ proteins. Molecular microbiology 2015 imex curation Osadnik H., Schöpfel M., Heidrich E., Mehner D., Lilie H., Parthier C., Risselada HJ., Grubmüller H., Stubbs MT., Brüser T. brueser@ifmb.uni-hannover.de Only protein-protein interactions imex curation in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Osadnik H., Schöpfel M., Heidrich E., Mehner D., Lilie H., Parthier C., Risselada HJ., Grubmüller H., Stubbs MT., Brüser T. brueser@ifmb.uni-hannover.de Accepted 2020-JUN-26 AT 14:30 BST AT 14:30 BST by LIVIAP Molecular microbiology 2015 Only protein-protein interactions imex curation imex curation Mock publication and experiment for abstract interactions that are not interaction evidences. Mock publication and experiment for abstract interactions that are not interaction evidences. unspecified method PspF-binding domain PspA1-144 and the PspA·F complex: New insights into the coiled-coil-dependent regulation of AAA+ proteins. PspF-binding domain PspA1-144 and the PspA·F complex: New insights into the coiled-coil-dependent regulation of AAA+ proteins. Molecular microbiology 2015 imex curation Osadnik H., Schöpfel M., Heidrich E., Mehner D., Lilie H., Parthier C., Risselada HJ., Grubmüller H., Stubbs MT., Brüser T. brueser@ifmb.uni-hannover.de Only protein-protein interactions imex curation in vitro In vitro solution sedimentati cosedimentation in solution solution sedimentati predetermined predetermined participant predetermined Osadnik H., Schöpfel M., Heidrich E., Mehner D., Lilie H., Parthier C., Risselada HJ., Grubmüller H., Stubbs MT., Brüser T. brueser@ifmb.uni-hannover.de Accepted 2020-JUN-26 AT 14:30 BST AT 14:30 BST by LIVIAP Molecular microbiology 2015 Only protein-protein interactions imex curation imex curation PspF-binding domain PspA1-144 and the PspA·F complex: New insights into the coiled-coil-dependent regulation of AAA+ proteins. PspF-binding domain PspA1-144 and the PspA·F complex: New insights into the coiled-coil-dependent regulation of AAA+ proteins. Molecular microbiology 2015 imex curation Osadnik H., Schöpfel M., Heidrich E., Mehner D., Lilie H., Parthier C., Risselada HJ., Grubmüller H., Stubbs MT., Brüser T. brueser@ifmb.uni-hannover.de Only protein-protein interactions imex curation in vitro In vitro atpase assay atpase assay predetermined predetermined participant predetermined Osadnik H., Schöpfel M., Heidrich E., Mehner D., Lilie H., Parthier C., Risselada HJ., Grubmüller H., Stubbs MT., Brüser T. brueser@ifmb.uni-hannover.de Accepted 2020-JUN-26 AT 14:30 BST AT 14:30 BST by LIVIAP Molecular microbiology 2015 Only protein-protein interactions imex curation imex curation PspF-binding domain PspA1-144 and the PspA·F complex: New insights into the coiled-coil-dependent regulation of AAA+ proteins. PspF-binding domain PspA1-144 and the PspA·F complex: New insights into the coiled-coil-dependent regulation of AAA+ proteins. Molecular microbiology 2015 imex curation Osadnik H., Schöpfel M., Heidrich E., Mehner D., Lilie H., Parthier C., Risselada HJ., Grubmüller H., Stubbs MT., Brüser T. brueser@ifmb.uni-hannover.de Only protein-protein interactions imex curation ecoli Escherichia coli (strain K12) anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Osadnik H., Schöpfel M., Heidrich E., Mehner D., Lilie H., Parthier C., Risselada HJ., Grubmüller H., Stubbs MT., Brüser T. brueser@ifmb.uni-hannover.de Accepted 2020-JUN-26 AT 14:30 BST AT 14:30 BST by LIVIAP Molecular microbiology 2015 Only protein-protein interactions imex curation imex curation atp ATP Adenosine 5'-triphosphate Adenosine triphosphate ADENOSINE-5'-TRIPHOSPHATE Adephos Adetol Adynol Atipi ATP Atriphos Cardenosine Fosfobion Glucobasin H4atp Myotriphos Triadenyl Triphosphaden adenosine 5'-(tetrahydrogen triphosphate) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) pspa_ecoli Phage shock protein A pspA b1304 JW1297 protein protein ecoli Escherichia coli (strain K12) MGIFSRFADIVNANINALLEKAEDPQKLVRLMIQEMEDTLVEVRSTSARALAEKKQLTRRIEQASAREVEWQEKAELALLKEREDLARAALIEKQKLTDLIKSLEHEVTLVDDTLARMKKEIGELENKLSETRARQQALMLRHQAANSSRDVRRQLDSGKLDEAMARFESFERRIDQMEAEAESHSFGKQKSLDDQFAELKADDAISEQLAQLKAKMKQDNQ EAC29290406E5053 pspf_ecoli Psp operon transcriptional activator pspF b1303 JW1296 ycjB Phage shock protein F protein protein ecoli Escherichia coli (strain K12) MAEYKDNLLGEANSFLEVLEQVSHLAPLDKPVLIIGERGTGKELIASRLHYLSSRWQGPFISLNCAALNENLLDSELFGHEAGAFTGAQKRHPGRFERADGGTLFLDELATAPMMVQEKLLRVIEYGELERVGGSQPLQVNVRLVCATNADLPAMVNEGTFRADLLDRLAFDVVQLPPLRERESDIMLMAEYFAIQMCREIKLPLFPGFTERARETLLNYRWPGNIRELKNVVERSVYRHGTSDYPLDDIIIDPFKRRPPEDAIAVSETTSLPTLPLDLREFQMQQEKELLQLSLQQGKYNQKRAAELLGLTYHQFRALLKKHQI F495D634193A42FB pspa-pspf-3 5 7 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P0AFM6:p.Glu37Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain E A Tue Jun 30 19:11:20 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu37Ala' 8 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed region strep tag strep tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. 4E, Supp FIg. 4E pspa-pspf-4 1 7 unspecified role unspecified role neutral component neutral component purified purified c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction Supp FIg. 5A pspa-pspf-5 3 7 unspecified role unspecified role neutral component neutral component purified purified c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P0AFM6:p.Arg59Glu mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain R E Tue Jun 30 19:55:02 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg59Glu' P0AFM6:p.Glu37Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain E A Tue Jun 30 20:17:02 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu37Ala' ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction FIg. 5A pspf-1 3 8 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction Supp FIg. 5B, 5D Stable oligomerization of PspF1-265 was not promoted by addition of ADP atp-pspaf_ecoli-1 4 35 enzyme enzyme unspecified role unspecified role purified purified c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 37 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 39 c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 39 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 37 ecoli Escherichia coli (strain K12) 6 enzyme target enzyme target substrate substrate unspecified role unspecified role atpase reaction atpase reaction Supp. Fig. 5E As they don't show pspF ATPase activity on its own, I have just kept this a simple reaction of the basal activity by the complex, and ignored the (variable) effect of the mutants. I don't think I can deal with those in any way. pspaf_ecoli pspAF transcription regulation complex 6xpspA:6xpspF 7 unspecified role unspecified role pspf binding region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain pspf binding region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 8 enzyme enzyme pspa binding region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain pspa binding region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 43 45 46 44 physical association physical association ecoli Escherichia coli (strain K12) stable complex stable complex physical interaction evidence used in manual assertion physical interaction evidence used in manual assertion IPI inferred from physical interaction Transcription regulator, formation of which in the absence of membrane stress inhibits the ATPase activity of the enhancer binding and activator protein pspF. ATPase activity of pspF is required to induce an open complex formation of the DNA‐bound RNA polymerase at its target promoters. The pspAF complex retains only a basal ATPase activity. Membrane stresses, such as the mislocalisation of proteins to the inner membrane, appear to lead to the recruitment of pspA to the pspBC complex (CPX-5892), enabling at least a partial release of pspA from pspF. This allows transcription of the Psp membrane-stabilizing system through binding of pspF to the sigma factor rpoN (P24255). pspA binds to the pspF hexamer and inhibits pspF activity by direct interaction of the AAA+ transcription activation domain of pspF. pspA appears to bind to Trp-56 of pspF. This is part of the 50-62 aa loop which contains a group of three amino acids, called the YLW patch, that serve as a docking site for pspA and for ATP hydrolytic activity. The C-terminal domain of pspA appears to be crucial for the regulation of pspF in response to membrane stress stimuli. Heterododecamer Accepted 2020-JUL-01 by LIVIAP