Role of the CCAAT-binding protein CBF/NF-Y in transcription

doi:10.1016/S0968-0004(98)01201-8

Trends in Biochemical Sciences

Volume 23, Issue 5, 1 May 1998, Pages 174-178

https://doi.org/10.1016/S0968-0004(98)01201-8 Get rights and content

Abstract

The CCAAT motif is one of the common promoter elements present in the proximal promoter of numerous mammalian genes transcribed by RNA polymerase II. CBF (also called NF-Y and CP1) consists of three different subunits and interacts specifically with the CCAAT motif. In each CBF subunit, the segment needed for formation of the CBF–DNA complex is conserved from yeast to human and, interestingly, the conserved segment of two CBF subunits, CBF-A and CBF-C, are homologous to the histone-fold motif of eukaryotic histones and archaebacterial histone-like protein HMf-2. The histone fold motifs of CBF-A and CBF-C interact with each other to form a heterodimer that associates with CBF-B to form a heterotrimeric CBF molecule, which then binds to DNA.

Section snippets

The CCAAT motif is a functional promoter element

Generally, mutations in the CCAAT motif cause a several-fold decrease in transcription activity both in vitro and in vivo3, 4, 5, 6. Table 1 shows examples of different mammalian promoters, which have been divided into three groups based on the effect of a CCAAT motif mutation on promoter activity. Mutations in the CCAAT motifs of group 1 promoters result in decreased transcription in the absence of stimulatory influences4, 5, 6, 7, 8, 9, 10. In promoters of group 2, similar mutations alter

CBF/NF-Y binds specifically to the CCAAT motif

To date, several proteins have been isolated that bind to CCAAT motifs; among these, CBF/NF-Y (also called CP1; referred to herein as CBF; see Appendix A) requires a high degree of conservation of the CCAAT sequence. Indeed, nucleotide-substitution mutations in each base of the CCAAT motif result in either loss or decrease in formation of the CBF–DNA complex. Other CCAAT-binding proteins include CTF/NF1 and CCAAT/enhancer-binding protein (C/EBP), but DNA sequences in the binding sites of these

Subunit assembly and DNA binding

Recombinant CBF polypeptides have been used to demonstrate that CBF-A and CBF-C interact with each other to form a stable heterodimer. CBF-B only interacts with the CBF-A/CBF-C heterodimer, not with CBF-A or CBF-C alone, to form a heterotrimeric CBF molecule, which then binds to DNA. The conserved segments of CBF-A and CBF-C that are necessary for formation of a CBF–DNA complex show amino acid sequence homology with each other. Interestingly, these two CBF segments display amino acid sequence

The flanking sequences of the CCAAT motif as determinants of CBF-binding affinity

Analysis of the CBF-binding sites indicates that mutations of each nucleotide of the CCAAT sequence in promoters decreases or abolishes CBF binding. Also, not all promoter DNAs containing the CCAAT sequence bind CBF, suggesting that specific nucleotide sequences other than CCAAT are necessary for CBF binding. Specific DNA sequences that are required for CBF binding have been defined by using PCR-mediated random binding selection[33]. This experiment indicated that the affinity of CBF binding to

Two separate transcriptional activation domains

Transcription of the α2(1) collagen promoter and of the Rous sarcoma virus long terminal repeat (RSV-LTR) is stimulated severalfold by the addition of purified recombinant CBF subunits to mouse nuclear extracts that were previously depleted of CBF. Analysis of CBF deletion mutants in such an in vitro transcription system identified two transcription activation domains in the heterotrimeric CBF, one present in CBF-B, the other in CBF-C, which activate transcription additively. The two CBF

Transcriptional regulation of different mammalian promoters by CBF

One way that CBF might regulate the transcription of various promoters is by cooperative interactions with other transcription factors that bind to a specific promoter. The liver-specific serum albumin promoter contains a CBF-binding site that is juxtaposed to a strong binding site for the C/EBP transcription factor. C/EBP and CBF activate transcription synergistically in liver nuclear extracts in vitro. However, this transcriptional synergism is not observed when the CBF binding site is moved

Concluding remarks and perspectives

CBF is a unique DNA-binding protein that interacts with the CCAAT motif, a common element present in various mammalian promoters. Portions of each of the three CBF subunits that are needed for DNA binding are conserved from yeast to human. The CCAAT motif is present in a limited number of yeast promoters and the CBF homolog in yeast regulates the transcription of nuclear genes that are involved in mitochondrial function. The conserved segments of two CBF subunits show homology with the ancient

Acknowledgements

Work performed in the laboratories of the authors was supported by grants from the National Institutes of Health.

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Trends in Biochemical Sciences

Role of the CCAAT-binding protein CBF/NF-Y in transcription

Abstract

Section snippets

The CCAAT motif is a functional promoter element

CBF/NF-Y binds specifically to the CCAAT motif

Subunit assembly and DNA binding

The flanking sequences of the CCAAT motif as determinants of CBF-binding affinity

Two separate transcriptional activation domains

Transcriptional regulation of different mammalian promoters by CBF

Concluding remarks and perspectives

Acknowledgements

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Cell

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Trends Genet.

Methods Enzymol.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Science