TY  - JOUR
A1  - Rogov, Vladimir V.
A1  - Rogova, Natalia Yu.
A1  - Bernhard, Frank
A1  - Löhr, Frank
A1  - Dötsch, Volker
T1  - A disulfide bridge network within the soluble periplasmic domain determines structure and function of the outer membrane protein RCSF
T2  - Journal of biological chemistry
N2  - RcsF, a proposed auxiliary regulator of the regulation of capsule synthesis (rcs) phosphorelay system, is a key element for understanding the RcsC-D-A/B signaling cascade, which is responsible for the regulation of more than 100 genes and is involved in cell division, motility, biofilm formation, and virulence. The RcsC-D-A/B system is one of the most complex bacterial signal transduction pathways, consisting of several membrane-bound and soluble proteins. RcsF is a lipoprotein attached to the outer membrane and plays an important role in activating the RcsC-d-A/B pathway. The exact mechanism of activation of the rcs phosphorelay by RcsF, however, remains unknown. We have analyzed the sequence of RcsF and identified three structural elements: 1) an N-terminal membrane-anchored helix (residues 3-13), 2) a loop (residues 14-48), and 3) a C-terminal folded domain (residues 49-134). We have determined the structure of this C-terminal domain and started to investigate its interaction with potential partners. Important features of its structure are two disulfide bridges between Cys-74 and Cys-118 and between Cys-109 and Cys-124. To evaluate the importance of this RcsF disulfide bridge network in vivo, we have examined the ability of the full-length protein and of specific Cys mutants to initiate the rcs signaling cascade. The results indicate that the Cys-74/Cys-118 and the Cys-109/Cys-124 residues correlate pairwise with the activity of RcsF. Interaction studies showed a weak interaction with an RNA hairpin. However, no interaction could be detected with reagents that are believed to activate the rcs phosphorelay, such as lysozyme, glucose, or Zn(2+) ions.
KW  - Bacterial Signal Transduction
KW  - Histidine Kinases
KW  - Lipoprotein Structure
KW  - Metabolic Regulation
KW  - NMR
KW  - NMR Solution Structure
KW  - Capsule Synthesis
KW  - Disulphide Bridges
KW  - Outer Membrane Protein RcsF
KW  - Two-component System
Y1  - 2021
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/76585
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-765858
SN  - 0021-9258
VL  - 286.2011
IS  - 21
SP  - 18775
EP  - 18783
PB  - American Society for Biochemistry and Molecular Biology Publications
CY  - Bethesda, Md
ER  -