Group leader: Bé Wieringa.

Group name: Department of Cell Biology, NCMLS UMCN.

Other Staff members: Jack Fransen (UHD), Ineke van der Zee (UD), Frank Oerlemans (technician), Helma Pluk (post-doc), Edwin Janssen (post-doc), Wieke de Bruin (post-doc), Jan Kuiper (Ph.D.student), Femke Streijger (Ph.D.student).

Location: NCMLS UMCN University Nijmegen.

Web page: www.ncmls.kun.nl

E.mail: b.wieringa@ncmls.kun.nl.

Phone: 31-24-3614329/3614287; Fax: 31-24-3615317.

Relevant research interests:

The cellular energetics network for high-energy phosphoryl (i.e. mainly ATP) transfer in mammalian cells consists of several redundant pathways, in which several glycolytic enzymes, members of the creatine kinase (CK) and adenylate kinase (AK) families of enzymes and nucleoside diphosphate kinases (NDPK) play a determining role. Reactions in this complex network are highly compartmentalized and require delicate coupling between cytosolic and organellar events, with coordinated control of cytoarchitectural arrangements and mitochondrial biogenesis and respiration, in a cell-type and developmental-stage dependent manner. It has now been well established that the individual enzymes in the ~P-transfer network work in close physical and functional association with the systems for cell motility and contraction (muscle, polarized migrating cells), intracellular transport and ion homeostasis (muscle, neural cells) machinery. Moreover, the same network also serves to protect cells against ischemic and anoxic stress and in providing (cancer) cells with specially endowed survival skills.

During the past decade our group has been employing reverse genetic approaches with gene knockout and pharmacological inhibition in cellular and animal models to study the cellular effects of intrinsic or extrinsic metabolic stress in the ~P transfer network in muscle, brain and cancer cells. Our studies with cDNA arrays, semiquantitative Western blotting, ~P flux measurements and physiological and behavioral analysis with biochemical, cell biological, NMR/MRI procedures have revealed that every pertubation in the large-scale organized energetics network (CK-AK knockout or glycolytic enzyme impairment) results in a cell- and mutation-type dependent multigene/multitranscript/mutiprotein response. Adaptation and remodeling involved (i) regulation at the transcriptional and post-transcriptional level, (ii) changes in the positioning and dynamics of the cytoarchitectural organization, and (iii) rewiring of activity through redundant ~P transfer pathways.

Current system biology activities:

Our present studies focus on the combined use of genomics-proteomics-and metabolomics approaches to unravel the control of regulatory metabolic-status signaling pathways (calcineurin-NFat, AMPK, mTOR and transcriptional co-activator events) that sense the metabolic stress in the ~P transfer network and contribute to the adaptational response. Moreover, we now concentrate on the study of the molecular environment of CK, AK and glycolytic enzymes and the use of quantitative real-time dynamic microscopy imaging methods to follow metabolite (ion) and enzyme behavior, to better understand their role in cellular energetics.

Relevant collaborations: Together with the group of dr. Peter Willems/W.Koopman (Dept. Biochemistry NCMLS, UMCN) we work on a computer modeling environment for quantitative description of changes in calcium-ion homeostasis, protein and organelle (mitochondrial)dynamics, and metabolite (NADH, ATP) distribution modes. At the international level we work together with dr. A.Terzic/P.Dzeja for ~P flux and physiological modeling.

Representative publications:

See www.ncmls.kun.nl dept. Cell Biology for background info and movies.

1. Deursen, J. van, Heerschap, A., Oerlemans, F., Ruitenbeek, W., Jap, P., Laak, H. ter, & Wieringa, B. (1993). Skeletal Muscles of Mice Deficient in Muscle Creatine Kinase Lack Burst Activity. Cell, 74, 621-631.

2. Deursen, J. van, Ruitenbeek, W., Heerschap, A., Jap, P., Laak, H. ter, & Wieringa, B (1994). Creatine kinase (CK) in skeletal muscle energy metabolism: A study of mouse mutants with graded reduction in muscle CK expression. Proc.Natl.Acad Sci USA 91, 9091-9095.

3. Steeghs, K., Benders, A., Oerlemans, F., de Haan, A., Heerschap, A., Ruitenbeek, W., Jost, C., van Deursen, J., Perryman, B., Pette, D., Brückwilder, M., Koudijs, J., Jap, P., Veerkamp, J. and Wieringa. B. (1997) Altered Ca²⁺ responses in muscles with combined mitochondrial and cytosolic creatine kinase deficiencies. Cell 89, 93-103.

4. Janssen, E., Dzeja, P.P., Oerlemans, F., Simonetti, A.W., Heerschap, A., de Haan, A., Rush, P.S., Terjung, R.R., Wieringa, B.and Terzic, A. (2000) Adenylate kinase 1 gene deletion disrupts muscle energetic economy despite metabolic rearrangement. EMBO J. 19, 6371-6381.

5. de Groof,AJ; Oerlemans,FT; Jost,CR; Wieringa,B. Changes in glycolytic network and mitochondrial design in creatine kinase-deficient muscles (2001) Muscle-Nerve. 24(9): 1188-96.

6. Jost, C., van der Zee, C.E.E.M., in ’t Zandt, H.J.A., Oerlemans, F., Verheij, M., Streijger, F., Fransen, J., van Deursen, J., Heerschap, A., Cools, A. and Wieringa, B. (2002) Creatine kinase B-driven energy transfer in the brain is important for habituation and spatial learning behaviour, mossy fibre field size and determination of seizure susceptibility. Eur. J. Neurosci. 15, 1692-1706.

7. Janssen, E., de Groof, A, Wijers M, Fransen, J., Dzeja P.P., Terzic A., and Wieringa, B. (2003). Adenylate kinase 1 deficiency induces molecular and structural adaptations to support muscle energy metabolism. J.Biol.Chem. 278, in press.