Structure Determination of TrmB and TrmB-like Transcriptional Regulators from the hyperthermophilic Archaeon Pyrococcus furiosus

• Applicant: Professor Dr. Wolfram Welte
• Subject Area: Structural Biology
• Term: from 2008 to 2013
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 91747536

TrmB is a novel transcriptional regulator of Pyrococcus furiosus that has dual effector and operator specificity. We propose to crystallize it with its different effectors: maltose, glucose and sucrose bound to it, as well as complexed with its two cognate operator sequences, the TM (Trehalose/Maltose) and the MD (Maltodextrin) operator sequence. This will reveal the two distinct modes of operator recognition as well as the differential effector response. The TrmB like regulator, TrmBLI, is a global regulator serving genes encoding glycolytic enzymes as represser and genes encoding gluconeogenic enzymes as activator. Here again, the structures of TrmBLI in complex with its effectors maltose, maltotriose and glucose and in complex with DMA harbouring the TGM (Thermococcales Glycolytic Motif) sequence of representative glycolytic and gluconeogenic genes will be determined. TrmBL2, the third homologous regulator lacks the effector binding site but is expected to interact with cellular proteins for transcriptional specificity. We plan to identify its interaction partner(s) and attempt to determine the structure of the complex.

• DFG Programme: Research Grants
• Participating Persons: Professor Dr. Winfried Boos; Professor Dr. Kay Diederichs