Identification of a functional CRE in the promoter of Fukuyama congenital muscular dystrophy gene fukutin.
We isolated a fragment of the fukutin gene promoter from differentiated human NT2 cells using chromatin immunoprecipitation technique with an anti-CREB antibody. This fragment contained a CRE-like sequence and here we describe its functional validation. The results showed that the element was functional in vitro and in vivo and that CREB in neurons was involved in the transcriptional regulation of the fukutin gene. Moreover, its expression in neurons was regulated by cAMP and calcium ions, known triggers of CREB phosphorylation. To our knowledge, this is the first report on the regulation of fukutin gene by transcription factor CREB in response to the signals generated by synaptic activity. The true biological function of fukutin, the gene responsible for Fukuyama-type congenital muscular dystrophy and mental retardation, is at present not known. However, it has been suggested that it might possess glycosyltransferase activity and its intracellular localization within the Golgi structures is consistent with this function. As such, fukutin might play a significant role in post-translational modification of synaptic proteins in neuronal cells.
Fang
Hung
H
Neurogenesis and Brain Repair Group, Institute for Biological Sciences, National Research Council of Canada, Ottawa, Ontario, Canada K1A 0R6. hung.fang@nrc.ca
Sodja
Caroline
C
Chartier
Joanne
J
Desbois
Angele
A
Lei
Joy
J
Walker
P Roy
PR
Sikorska
Marianna
M
D001324:Autoantigens / Q000378:metabolism; D015152:Blotting, Northern / Q000379:methods; D015153:Blotting, Western / Q000379:methods; D045744:Cell Line, Tumor; D047369:Chromatin Immunoprecipitation / Q000379:methods; D003001:Cloning, Molecular; D005576:Colforsin / Q000494:pharmacology; D017362:Cyclic AMP Response Element-Binding Protein / Q000302:isolation & purification* / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay / Q000379:methods; D005455:Fluorescent Antibody Technique / Q000379:methods; D005786:Gene Expression Regulation / Q000187:drug effects; D017930:Genes, Reporter / Q000502:physiology; D000077614:Golgi Matrix Proteins; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins; D011189:Potassium Chloride / Q000494:pharmacology; D011401:Promoter Regions, Genetic / Q000502:physiology*; D011485:Protein Binding; D011506:Proteins / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013347:Subcellular Fractions / Q000378:metabolism; D018243:Teratocarcinoma; D015533:Transcriptional Activation / Q000502:physiology; D014162:Transfection / Q000379:methods
D001324:Autoantigens; D017362:Cyclic AMP Response Element-Binding Protein; C113587:FKTN protein, human; D000077614:Golgi Matrix Proteins; C450721:Golgi complex autoantigen, 97-kDa; D008565:Membrane Proteins; D011506:Proteins; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins; D005576:Colforsin; D011189:Potassium Chloride