Brigham Young University BYU ScholarsArchive All Theses and Dissertations 2013-03-15 Evidences for Protein-Protein Interactions Between PstB and PhoU in the Phosphate Signaling Complex of Escherichia coli Kristine Dawn Johns Brigham Young University - Provo Follow this and additional works at: http://scholarsarchive.byu.edu/etd Part of the Microbiology Commons BYU ScholarsArchive Citation Johns, Kristine Dawn, "Evidences for Protein-Protein Interactions Between PstB and PhoU in the Phosphate Signaling Complex of Escherichia coli" (2013). All Theses and Dissertations. Paper 3932. This Thesis is brought to you for free and open access by BYU ScholarsArchive. It has been accepted for inclusion in All Theses and Dissertations by an authorized administrator of BYU ScholarsArchive. For more information, please contact [email protected]. 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SURSRVHWKDWGLUHFWSURWHLQSURWHLQLQWHUDFWLRQVEHWZHHQ3KR8DQGWKHDOWHUQDWLQJFRQIRUPDWLRQV RI3VW6&$%PHGLDWH3KR8LQWHUDFWLRQVZLWK3KR5%\PHDQVRIJHQHWLFDQGELRFKHPLFDO DSSURDFKHVZHKDYHIRXQGVXEVWDQWLDOHYLGHQFHVXSSRUWLQJERWKWKHVHK\SRWKHVHV .H\ZRUGV3KRUHJXORQWZRFRPSRQHQW$%&WUDQVSRUWHU3VW%3KR8 TABLE OF CONTENTS TITLE PAGE««««««««««««««««««««««««««««««««««««««««««««««««««i ABSTRACT«««««««««««««««««««««««««««««««««««««««««««««««.««««ii TABLE OF CONTENTS ............................................................................................................................................ iii LIST OF TABLES......................................................................................................................................................... v LIST OF FIGURES ......................................................................................................................................................vi INTRODUCTION ......................................................................................................................................................... 1 Two-component signaling .........................................................................................................................................2 Histidine Kinases ....................................................................................................................................................... 2 Extracellular domain ..............................................................................................................................................3 Cytoplasmic domain ..............................................................................................................................................4 Response Regulators ..................................................................................................................................................5 REC domain .......................................................................................................................................................... 5 Effector domain ..................................................................................................................................................... 6 ABC Transporters ...................................................................................................................................................... 7 Transmembrane domain ........................................................................................................................................8 Nucleotide-binding domain ...................................................................................................................................8 Conformational changes ........................................................................................................................................9 The Pho Regulon ....................................................................................................................................................... 9 The two-component system, PhoR-PhoB ................................................................................................................ 10 PhoB ± Response Regulator (RR) ....................................................................................................................... 10 PhoR ± Sensory Histidine Kinase (HK)............................................................................................................... 11 The ABC transporter, PstSCAB2 ............................................................................................................................. 12 TMD- PstA and PstC ........................................................................................................................................... 12 NBD- PstB dimer................................................................................................................................................. 13 PhoU involvement in signal transduction ................................................................................................................ 13 MATERIALS AND METHODS ................................................................................................................................ 16 iii 0HGLDDQGJURZWKFRQGLWLRQV %DFWHULDOVWUDLQFRQVWUXFWLRQ 3ODVPLGFRQVWUXFWLRQ 0XWDQWFRQVWUXFWLRQ '1$HOHFWURSKRUHVLV 7UDQVIRUPDWLRQV $ONDOLQHSKRVSKDWDVHDVVD\V 1LFNHOFROXPQFKURPDWRJUDSK\ :HVWHUQEORWWLQJ %DFWHULDOWZRK\EULGVFUHHQ ȕJDODFWRVLGDVHDVVD\V 3KRVSKDWHWUDQVSRUWDVVD\ 5(68/76 $LP,GHQWLI\GLUHFWSURWHLQSURWHLQLQWHUDFWLRQVEHWZHHQ3VW%DQG3KR8 'HWHUPLQHIXQFWLRQDOLW\RIUHFRQVWLWXWHG3VW%DQG3VW%+LV (VWDEOLVKGRPLQDQFHRIFKURPRVRPDOSVW%DQGSODVPLGEDVHGSVW%H[SUHVVLRQ 3VW%3KR8LQWHUDFWLRQVREVHUYHGYLDFRSXULILFDWLRQDQGLPPXQREORWWLQJ 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