Dissection of melanogenesis with small molecules identifies prohibitin as a regulator.

Bioactive compounds can be used to selectively modulate gene function. We utilized a chemical genetic approach to dissect the mammalian pigmentation pathway and identify protein regulators. We screened a tagged library of 1170 small molecules in a cell-based assay and discovered a class of pigment-enhancing chemicals. From this class we characterized the small molecule melanogenin. Using melanogenin bound to an affinity matrix and amino acid sequencing, we identified the mitochondrial protein, prohibitin, as an intracellular binding target. Studies employing siRNA demonstrate that prohibitin is required for melanogenin to exert its propigmentary effects and reveal an unsuspected functional role for this protein in melanin induction. This represents a mechanism by which propigmentary signals are transduced and ultimately provides a potential target for the treatment of pigmentary disorders.

[1]  M. Schmitz,et al.  Prohibitin and prohibitone are contained in high-molecular weight complexes and interact with alpha-actinin and annexin A2. , 2002, Biochimie.

[2]  J. Schneider-Mergener,et al.  Spatially addressed synthesis of amino- and amino-oxy-substituted 1, 3,5-triazine arrays on polymeric membranes. , 2000, Journal of combinatorial chemistry.

[3]  S. Schreiber,et al.  SIR1, an Upstream Component in Auxin Signaling Identified by Chemical Genetics , 2003, Science.

[4]  P. Jat,et al.  BAP37 and Prohibitin are specifically recognized by an SV40 T antigen antibody. , 2000, Molecular cell biology research communications : MCBRC.

[5]  S. Chellappan,et al.  Prohibitin Induces the Transcriptional Activity of p53 and Is Exported from the Nucleus upon Apoptotic Signaling* , 2003, Journal of Biological Chemistry.

[6]  L. Burdine,et al.  Target identification in chemical genetics: the (often) missing link. , 2004, Chemistry & biology.

[7]  D. Faller,et al.  Prohibitin requires Brg‐1 and Brm for the repression of E2F and cell growth , 2002, The EMBO journal.

[8]  D. Bennett,et al.  Cloned mouse melanocyte lines carrying the germline mutations albino and brown: complementation in culture. , 1989, Development.

[9]  E. Jupe,et al.  Prohibitin: Potential role in senescence, development, and tumor suppression , 1995, Experimental Gerontology.

[10]  S. Pomerantz.  The tyrosine hydroxylase activity of mammalian tyrosinase. , 1966, The Journal of biological chemistry.

[11]  L. Grivell,et al.  The mitochondrial PHB complex: roles in mitochondrial respiratory complex assembly, ageing and degenerative disease , 2002, Cellular and Molecular Life Sciences CMLS.

[12]  R. Buscà,et al.  Microphthalmia-associated transcription factor (MITF) is required but is not sufficient to induce the expression of melanogenic genes. , 2003, Pigment cell research.

[13]  D. A. Brown,et al.  Skin pigmentation enhancers. , 2001, Journal of photochemistry and photobiology. B, Biology.

[14]  S. Orlow,et al.  Inhibition of induced melanogenesis in Cloudman melanoma cells by four phenotypic modifiers. , 1990, Experimental cell research.

[15]  L. Grivell,et al.  Prohibitins act as a membrane‐bound chaperone for the stabilization of mitochondrial proteins , 2000, The EMBO journal.

[16]  P. Hall,et al.  Mammalian prohibitin proteins respond to mitochondrial stress and decrease during cellular senescence. , 2001, Experimental cell research.

[17]  Da-Woon Jung,et al.  Identification of compounds that bind mitochondrial F1F0 ATPase by screening a triazine library for correction of albinism. , 2004, Chemistry & biology.

[18]  Derek S. Tan.  Sweet surrender to chemical genetics , 2002, Nature Biotechnology.

[19]  Tae-Wook Kang,et al.  Facilitated forward chemical genetics using a tagged triazine library and zebrafish embryo screening. , 2003, Journal of the American Chemical Society.

[20]  Chun-Ming Huang,et al.  Comparative proteomic profiling of murine skin. , 2003, The Journal of investigative dermatology.

[21]  S. Orlow,et al.  Accumulation of tyrosinase in the endolysosomal compartment is induced by U18666A. , 2003, Pigment cell research.

[22]  N. Nath,et al.  Prohibitin, a potential tumor suppressor, interacts with RB and regulates E2F function , 1999, Oncogene.

[23]  Makoto Kinoshita,et al.  [Small molecule inhibitor of mitotic spindle bipolarity identified in a phenotype-based screen]. , 2007, Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme.

[24]  D. Bennett,et al.  A line of non‐tumorigenic mouse melanocytes, syngeneic with the B16 melanoma and requiring a tumour promoter for growth , 1987, International journal of cancer.

[25]  Stuart L. Schreiber,et al.  Dissecting glucose signalling with diversity-oriented synthesis and small-molecule microarrays , 2002, Nature.

[26]  J Mottram,et al.  Intracellular targets of cyclin-dependent kinase inhibitors: identification by affinity chromatography using immobilised inhibitors. , 2000, Chemistry & biology.

[27]  A. Menssen,et al.  Characterization of the c-MYC-regulated transcriptome by SAGE: Identification and analysis of c-MYC target genes , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[28]  S. Schreiber,et al.  Target-oriented and diversity-oriented organic synthesis in drug discovery. , 2000, Science.

[29]  M. Staňková,et al.  Library generation through successive substitution of trichlorotriazine , 1996, Molecular Diversity.

[30]  Peter G. Schultz,et al.  Myoseverin, a microtubule-binding molecule with novel cellular effects , 2000, Nature Biotechnology.

[31]  Niharika Nath,et al.  Rb and Prohibitin Target Distinct Regions of E2F1 for Repression and Respond to Different Upstream Signals , 1999, Molecular and Cellular Biology.