CAP2, cyclase-associated protein 2, is a dual compartment protein.

Cyclase-associated proteins (CAPs) are evolutionarily conserved proteins with roles in regulating the actin cytoskeleton and in signal transduction. Mammals have two CAP genes encoding the related CAP1 and CAP2. We studied the distribution and subcellular localization of CAP1 and CAP2 using specific antibodies. CAP1 shows a broad tissue distribution, whereas CAP2 is significantly expressed only in brain, heart and skeletal muscle, and skin. CAP2 is found in the nucleus in undifferentiated myoblasts and at the M-line of differentiated myotubes. In PAM212, a mouse keratinocyte cell line, CAP2 is enriched in the nucleus, and sparse in the cytosol. By contrast, CAP1 localizes to the cytoplasm in PAM212 cells. In human skin, CAP2 is present in all living layers of the epidermis localizing to the nuclei and the cell periphery. In in vitro studies, a C-terminal fragment of CAP2 interacts with actin, indicating that CAP2 has the capacity to bind to actin.

Transcriptional repression by the zinc finger protein REST is mediated by titratable nuclear factors.

The zinc finger protein REST (RE-1 silencing transcription factor) is a transcriptional repressor that inhibits neuronal gene transcription in non-neuronal tissues. REST may represent a master regulator of neuronal gene expression. REST contains two repressor domains located at the N- and C-termini of the molecule. To investigate the molecular mechanism of transcriptional repression by REST, in vivo competition experiments were performed. Both repression domains were expressed in the nucleus as fusion proteins with S. japonicum glutathione S-transferase (GST). The ability of these fusion proteins to block transcriptional repression mediated by the repressor domains of REST was tested. The results show that transcriptional repression by the N-terminal repression domain of REST could be overcome by expression of a GST fusion protein encoding the N-terminal, but not C-terminal repression domain, and vice versa, suggesting that both repression domains have to interact with distinct nuclear factors to exhibit biological activity. The GST-REST fusion proteins had no effect upon transcriptional repression mediated by the KRAB (Krüppel-associated box) domain, a strong mammalian repressor domain, or the repressor domain derived from the thyroid hormone receptors alpha. We conclude that REST has to interact with at least two distinct nuclear factors to inhibit transcription. These factors are distinct from the mammalian corepressor proteins KAP-1/KRIP-1 and N-CoR that mediate repression by the KRAB domain or the thyroid hormone receptor alpha. Thus, mammalian transcriptional repressors utilize different mechanisms to inhibit transcription by using different kinds of protein-protein interactions.

Regulation of neuronal gene expression.

Humans as multicellular organisms contain a variety of different cell types where each cell population must fulfill a distinct function in the interest of the whole organism. The molecular basis for the variations in morphology, biochemistry, molecular biology, and function of the various cell types is the cell-type specific expression of genes. These genes encode proteins necessary for executing the specialized functions of each cell type within an organism. We describe here a regulatory mechanism for the expression of neuronal genes. The zinc finger protein REST binds to the regulatory region of many neuronal genes and represses neuronal gene expression in nonneuronal tissues. A negative regulatory mechanism, involving a transcriptional repressor, seems to play an important role in establishing the neuronal phenotype.