The core dimerization domains of histidine kinases contain recognition specificity for the cognate response regulator.
Histidine kinases DivJ and PleC initiate signal transduction pathways that regulate an early cell division cycle step and the gain of motility later in the Caulobacter crescentus cell cycle, respectively. The essential single-domain response regulator DivK functions downstream of these kinases to catalyze phosphotransfer from DivJ and PleC. We have used a yeast two-hybrid screen to investigate the molecular basis of DivJ and PleC interaction with DivK and to identify other His-Asp signal transduction proteins that interact with DivK. The only His-Asp proteins identified in the two-hybrid screen were five members of the histidine kinase superfamily. The finding that most of the kinase clones isolated correspond to either DivJ or PleC supports the previous conclusion that DivJ and PleC are cognate DivK kinases. A 66-amino-acid sequence common to all cloned DivJ and PleC fragments contains the conserved helix 1, helix 2 sequence that forms a four-helix bundle in histidine kinases required for dimerization, autophosphorylation and phosphotransfer. We present results that indicate that the four-helix bundle subdomain is not only necessary for binding of the response regulator but also sufficient for in vivo recognition specificity between DivK and its cognate histidine kinases. The other three kinases identified in this study correspond to DivL, an essential tyrosine kinase belonging to the same kinase subfamily as DivJ and PleC, and the two previously uncharacterized, soluble histidine kinases CckN and CckO. We discuss the significance of these results as they relate to kinase response regulator recognition specificity and the fidelity of phosphotransfer in signal transduction pathways.
Ohta
Noriko
N
Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544, USA.
Newton
Austin
A
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000378:metabolism*; D016935:Caulobacter crescentus / Q000737:chemistry / Q000201:enzymology* / Q000378:metabolism; D019281:Dimerization; D015964:Gene Expression Regulation, Bacterial; D000071677:Histidine Kinase; D008969:Molecular Sequence Data; D011494:Protein Kinases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D020798:Two-Hybrid System Techniques
D001426:Bacterial Proteins; C095478:DivK protein, Caulobacter crescentus; D011494:Protein Kinases; D000071677:Histidine Kinase