Solution structure of the dimeric SAM domain of MAPKKK Ste11 and its interactions with the adaptor protein Ste50 from the budding yeast: Implications for Ste11 activation and signal transmission through the Ste50-Ste11 complex

Ste11, a homologue of mammalian MAPKKKs, together with its binding partner Ste,50 works in a number of MAPK signaling pathways of Saccharomyces cerevisiae. Ste11/Ste50 binding is mediated by their sterile a motifs or SAM domains, of which homologues are also found in many other intracellular signaling and regulatory proteins. Here, we present the solution structure of the SAM domain or residues D37-R104 of Stell. and its interactions with the cognate SAM domain-containing region of Ste50, residues M27-Q131. NMR pulse-field-gradient (PFG) and rotational correlation time measurements (tau(c)) establish that the Stell SAM domain exists predominantly as a symmetric dimer in solution. The solution structure of the dimeric Stell SAM domain consists of five well-defined helices per monomer packed into a compact globular structure. The dimeric structure of the SAM domain is maintained by a novel dimer interface involving interactions between a number of hydrophobic residues situated on helix 4 and at the beginning of the C-terminal long helix (helix 5). The dimer structure may also be stabilized by potential salt bridge interactions across the interface. NMR H/H-2 exchange experiments showed that binding of the Ste50 SAM to the Stell SAM very likely involves the positively charged extreme C-terminal region as well as exposed hydrophobic patches of the dimeric Stell SAM domain. The dimeric structure of the Stell SAM and its interactions with the Ste50 SAM may have important roles in the regulation and activation of the Stell kinase and signal transmission and amplifications through the Ste50-Ste11 complex. Crown Copyright (C) 2004 Published by Elsevier Ltd. All rights reserved.