Adipocytes play a critical role in energy balance and metabolism in metazoan organisms. From analysis of
genetically altered mice and cultured cell lines, key transcription factors and cellular signaling steps in adipocyte
differentiation have been described. However, detailed insight into adipogenic mechanisms is still lacking.
A recently discovered NE-protein interacts with important lipogenic proteins, raising the possibility that it plays an
important role in adipogenesis.
Nuclear envelope(NE) proteins have been implicated in many human genetic diseases.
Therefore, I state a core hypothesis for this project proposal: "A specific NE-protein is required for
adipogenesis, by regulating functions of essential factors in adipogenesis".
To test this hypothesis I will set out to accomplish the following three aims:
Aim 1) To define the NE-protein`s role during adipogenic differentiation
In my core hypothesis I propose that the NE-protein is required for adipose cell differentiation. To test this it will
be silenced in 3T3L1 and OP9 cells by RNAi, and subsequently the effects on adipose cell differentiation will be
monitored.
Aim 2) To elucidate the NE-protein`s sub-cellular localization determinants and its possible interaction with
adipogenic factors
If the NE-protein is found to be essential for adipocyte development, its subcellular localization and its intrinsic
localization determinants will be studied by mutational analysis.
Moreover, the question if it physically interacts with adipogenic factors will be addressed using co-
immunoprecipitation.
Aim 3) To find out if the NE-protein and its specific activity are required for localization/activity of
adipogenic factors and/or adipose cell differentiation
If the NE-protein is found to be essential for adipogenesis and/or adipogenic factors` regulation mutational analysis
will be carried out to clarify if an intact active domain of the the NE-protein is essential for adipocyte
development.