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M402102200v1
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Papers In Press, published online ahead of print April 8, 2004
J. Biol. Chem, 10.1074/jbc.M402102200
Submitted on February 25, 2004
Revised on April 7, 2004
Accepted on April 8, 2004

Sequestration of thermogenic transcription factors in the cytoplasm during development of brown adipose tissue

Jong S. Rim, Bingzhong Xue, Barbara Gawronska-Kozak, and Leslie P. Kozak

Pennington Biomedical Research Center, Baton Rouge, LA 70808

Corresponding Author: kozaklp{at}pbrc.edu

ABSTRACT Transcription factors that regulate gene expression during adipogenesis also control the expression of genes of thermogenesis in brown adipose tissue, in particular, the mitochondrial uncoupling protein gene (Ucp1). There is evidence that a plasticity exists among adipocytes in which activation of the Ucp1 gene together with mitochondrial biogenesis can increase the brown adipocyte character of white fat. To understand this process we have characterized the changes in transcription that occur in interscapular brown adipocytes during development. We have found dramatic reductions in both DNA binding activity to probes and immunoreactive protein for PPAR, RXR, C/EBPs and CREB regulatory motifs in nuclear extracts when mice reach adulthood. Exposure of adult mice to the cold, which reactivates Ucp1 expression, leads to a re-accumulation of factors in the nucleus. We propose that transcription factors are sequestered in the cytoplasm as mice age under conditions of reduced thermogenesis. Changes in isoform sub-types for PPARg and CREBs indicates an additional level of control on gene expression during thermogenesis. The increased movement of the RIIb PKA regulatory subunit into the nucleus with age suggests a mechanism for regulating the phosphorylation of transcription factors in the nucleus in response to the thermogenic requirements of the animal. NF-kB has been used as a model to demonstrate that the nuclear localization of transcription factors in brown fat are reduced during post-natal developmental. Furthermore, it is found by immunofluoresence that adrenergic stimulation of primary adipocyte cultures causes an increase of both the PKAa catalytic subunit and NF-kB into the nucleus.


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