title: Interactions among HAMP domain repeats act as an osmosensing molecular switch in group III hybrid histidine kinases from fungi.
abstract: The members of group III hybrid histidine kinases (HHK) are ubiquitous in fungi. Group III HHK have been implicated to function as osmosensors in the high osmolarity glycerol (HOG) pathway that is essential for fungal survival under high osmolarity stress. Recent literature suggests that group III HHK are also involved in conidia formation, virulence in several filamentous fungi, and are an excellent molecular target for antifungal agents. Thus, group III HHK constitute a very important group of sensor kinases. Structurally, group III HHK are distinct from Sln1p, the osmosensing HHK that regulates the HOG pathway in Saccharomyces cerevisiae. Group III HHK lack any transmembrane domain and typically contain HAMP domain repeats at the N terminus. Until now, it is not clear how group III HHK function as an osmosensor to regulate the HOG pathway. To investigate this, we undertook molecular characterization of DhNIK1, an ortholog from osmotolerant yeast Debaryomyces hansenii. We show here that DhNIK1 could complement sln1 mutation in S. cerevisiae thereby confirming its role as a bona fide osmosensor. We further investigated the role of HAMP domains by deleting them systematically. Our results clearly indicate that the HAMP4 domain is crucial for osmosensing by DhNik1p. Most importantly, we also show that the alternative interaction among the HAMP domains regulates the activity of DhNik1p like an "on-off switch" and thus provides, for the first time, an insight into the molecular mechanism of osmosensing by this group of HHKs.
authors.affiliation: Institute of Microbial Technology, Council of Scientific and Industrial Research, Chandigarh, India.
authors.forename: Netrapal
authors.initials: N
authors.lastname: Meena
authors.forename: Harsimran
authors.initials: H
authors.lastname: Kaur
authors.forename: Alok K
authors.initials: AK
authors.lastname: Mondal
mesh_terms: D000595:Amino Acid Sequence; D055165:Debaryomyces / Q000201:enzymology / Q000235:genetics; D005658:Fungi / Q000201:enzymology* / Q000235:genetics; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D000071677:Histidine Kinase; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D009994:Osmolar Concentration; D010802:Phylogeny; D011494:Protein Kinases / Q000737:chemistry* / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020449:Repetitive Sequences, Amino Acid; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid; D020798:Two-Hybrid System Techniques
chemical_list: D011994:Recombinant Proteins; D011494:Protein Kinases; D000071677:Histidine Kinase