Transcription factor CP2 is essential for lens-specific expression of the chicken alphaA-crystallin gene.
BACKGROUND
Lens-specific transcriptional activation of the chicken alphaA-crystallin gene is controlled by the distal and proximal enhancers, alphaCE1 and alphaCE2, respectively. Analysis using specific monoclonal antibodies against purified alphaCE1-binding factor alphaCEF1 revealed that alphaCEF1 is composed of two distinct subunits.


RESULTS
We have demonstrated that one of the subunits of alphaCEF1 is encoded by chicken ubiquitous transcription factor CP2 (cCP2), which is homologous to mouse CP2, and human CP2/LBP-1/LSF-1. Electrophoretic mobility shift assays and cross-linking experiments showed that alphaCEF1 and bacterially expressed cCP2 form a tetramer. Overexpression of cCP2 activates transcription through alphaCE1, but a mutant cCP2 lacking the DNA-binding domain reduced the transcription to basal levels. Although cCP2 binds to the CP2 template from the mouse alpha-globin promoter, it fails to promote transcription through this template. Element substitution experiments between alphaCE1 and the CP2 template revealed that the lens-specific enhancer activity of alphaCE1 is due to the 6 bp sequence (-139/-134; lens-specific element (LSE)) adjacent to the 3' of the cCP2 binding site within alphaCE1.


CONCLUSIONS
We have shown that the tetrameric transcription factor cCP2 is essential for lens-specific transcription of the chicken alphaA-crystallin gene, although it is ubiquitously expressed. We propose a model where cCP2 cooperates with a putative lens-specific factor which binds to LSE.
Murata
T
T
Department of Biophysics, Graduate School of Science, Kyoto University, Japan.
Nitta
M
M
Yasuda
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002467:Cell Nucleus / Q000737:chemistry; D002642:Chick Embryo; D002645:Chickens; D003001:Cloning, Molecular; D003459:Crystallins / Q000235:genetics*; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004742:Enhancer Elements, Genetic / Q000235:genetics; D007908:Lens, Crystalline / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009928:Organ Specificity; D011487:Protein Conformation; D012333:RNA, Messenger / Q000032:analysis; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D017422:Sequence Analysis, DNA; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic / Q000235:genetics; D015533:Transcriptional Activation / Q000235:genetics*
D003459:Crystallins; D004268:DNA-Binding Proteins; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; C501906:TFCP2 protein, human; C501905:Tfcp2 protein, mouse; D014157:Transcription Factors; D004247:DNA