Cloning of murine TCF-1, a T cell-specific transcription factor interacting with functional motifs in the CD3-epsilon and T cell receptor alpha enhancers

CD3-epsilon gene expression is confined to the T cell lineage. We have recently identified and cloned a human transcription factor, TCF-1, that binds to a functional element in the T lymphocyte-specific enhancer of CD3-epsilon. In a panel of human cell lines, TCF-1 expression was restricted to T lineage cells. TCF-1 belonged to a novel family of genes that contain the so-called high mobility group 1 (HMG) box. Here we report the cloning of murine TCF-1. Two splice alternatives were identified that were not previously observed in human TCF-1. Murine and human TCF-1 displayed a 95.5% overall amino acid homology. Recombinant murine and human TCF-1 recognized the same sequence motif in the CD3-epsilon enhancer as judged by gel retardation and methylation interference assays. With the murine cDNA clones several aspects of TCF-1 were analyzed. First, deletion analysis revealed that a region of TCF-1 containing the HMG box was sufficient for sequence-specific binding. Second, by high stringency Northern blotting and in situ hybridization, TCF-1 expression was shown to be confined to the thymus and to the T cell areas of the spleen. Third, TCF-1 bound specifically to a functional T cell-specific element in the T cell receptor alpha (TCR-alpha) enhancer. The T lineage-specific expression and the affinity for functional motifs in the TCR-alpha and CD3-epsilon enhancers imply an important role for TCF-1 in the establishment of the mature T cell phenotype.

[1]  G. Felsenfeld,et al.  The erythroid-specific transcription factor eryf1: A new finger protein , 1989, Cell.

[2]  K. Jones,et al.  Purification of TCF-1 alpha, a T-cell-specific transcription factor that activates the T-cell receptor C alpha gene enhancer in a context-dependent manner. , 1990, The New biologist.

[3]  Robin Lovell-Badge,et al.  A gene from the human sex-determining region encodes a protein with homology to a conserved DNA-binding motif , 1990, Nature.

[4]  M. Rosenfeld,et al.  A tissue-specific transcription factor containing a homeodomain specifies a pituitary phenotype , 1988, Cell.

[5]  H. Clevers,et al.  Identification and cloning of TCF‐1, a T lymphocyte‐specific transcription factor containing a sequence‐specific HMG box. , 1991, The EMBO journal.

[6]  R. Roeder,et al.  A human lymphoid- specific transcription factor that activates immunoglobulin genes is a homoeobox protein , 1988, Nature.

[7]  E. A. O'neill,et al.  The proline-rich transcriptional activator of CTF/NF-I is distinct from the replication and DNA binding domain , 1989, Cell.

[8]  C Terhorst,et al.  Human CD3-epsilon gene contains three miniexons and is transcribed from a non-TATA promoter. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[9]  W. Schaffner,et al.  A cloned octamer transcription factor stimulates transcription from lymphoid–specific promoters in non–B cells , 1988, Nature.

[10]  M. Smith,et al.  Four mating‐type genes control sexual differentiation in the fission yeast. , 1988, The EMBO journal.

[11]  C Yanofsky,et al.  Neurospora crassa a mating-type region. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[12]  T. Maniatis,et al.  Regulation of inducible and tissue-specific gene expression. , 1987, Science.

[13]  H. Clevers,et al.  An enhancer located in a CpG‐island 3′ to the TCR/CD3‐epsilon gene confers T lymphocyte‐specificity to its promoter. , 1989, The EMBO journal.

[14]  D. Baltimore,et al.  A novel, inducible and T cell‐specific enhancer located at the 3′ end of the T cell receptor alpha locus. , 1989, The EMBO journal.

[15]  H. Weintraub,et al.  Expression of a single transfected cDNA converts fibroblasts to myoblasts , 1987, Cell.

[16]  S. McKnight,et al.  Eukaryotic transcriptional regulatory proteins. , 1989, Annual review of biochemistry.

[17]  C Kress,et al.  Hox-2.3 upstream sequences mediate lacZ expression in intermediate mesoderm derivatives of transgenic mice. , 1990, Development.

[18]  H C Clevers,et al.  Towards a molecular understanding of T-cell differentiation. , 1991, Immunology today.

[19]  I. Ho,et al.  The T alpha 2 nuclear protein binding site from the human T cell receptor alpha enhancer functions as both a T cell-specific transcriptional activator and repressor , 1990, The Journal of experimental medicine.

[20]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[21]  T. Deerinck,et al.  The pituitary-specific transcription factor GHF-1 is a homeobox-containing protein , 1988, Cell.

[22]  B Guss,et al.  Gene fusion vectors based on the gene for staphylococcal protein A. , 1983, Gene.

[23]  D G Wilkinson,et al.  A molecular analysis of mouse development from 8 to 10 days post coitum detects changes only in embryonic globin expression. , 1987, Development.

[24]  Robert Tjian,et al.  Nucleolar transcription factor hUBF contains a DNA-binding motif with homology to HMG proteins , 1990, Nature.

[25]  D. Lockshon,et al.  MyoD is a sequence-specific DNA binding protein requiring a region of myc homology to bind to the muscle creatine kinase enhancer , 1989, Cell.

[26]  Shih-Feng Tsai,et al.  Cloning of cDNA for the major DNA-binding protein of the erythroid lineage through expression in mammalian cells , 1989, Nature.

[27]  M. Atchison,et al.  Enhancers: mechanisms of action and cell specificity. , 1988, Annual Review of Cell Biology.