Role of the multifunctional CDP/Cut/Cux homeodomain transcription factor in regulating differentiation, cell growth and development

doi:10.1016/S0378-1119(01)00485-1

Gene

Volume 270, Issues 1–2, 30 May 2001, Pages 1-15

https://doi.org/10.1016/S0378-1119(01)00485-1 Get rights and content

Abstract

CDP/Cux/Cut proteins are an evolutionarily conserved family of proteins containing several DNA binding domains: one Cut homeodomain and one, two or three Cut repeats. In Drosophila melanogaster, genetic studies indicated that Cut functions as a determinant of cell-type specification in several tissues, notably in the peripheral nervous system, the wing margin and the Malpighian tubule. Moreover, Cut was found to be a target and an effector of the Notch signaling pathway. In vertebrates, the same functions appear to be fulfilled by two cut-related genes with distinct patterns of expression. Cloning of the cDNA for the CCAAT-displacement protein (CDP) revealed that it was the human homologue of Drosophila Cut. CDP was later found be the DNA binding protein of the previously characterized histone nuclear factor D (HiNF-D). CDP and its mouse counterpart, Cux, were also reported to interact with regulatory elements from a large number of genes, including matrix attachment regions (MARs). CDP/Cut proteins were found generally to function as transcriptional repressors, although a participation in transcriptional activation is suggested by some data. Repression by CDP/Cut involves competition for binding site occupancy and active repression via the recruitment of a histone deacetylase activity. Various combinations of Cut repeats and the Cut homeodomains can generate distinct DNA binding activities. These activities are elevated in proliferating cells and decrease during terminal differentiation. One activity, involving the Cut homeodomain, is upregulated in S phase. CDP/Cut function is regulated by several post-translational modification events including phosphorylation, dephosphorylation, and acetylation. The CUTL1 gene in human was mapped to 7q22, a chromosomal region that is frequently rearranged in various cancers.

Section snippets

CDP/Cut is conserved among metazoans

The CDP/Cut family constitutes a unique group of homeoproteins, conserved among higher eukaryotes, and containing a Cut homeodomain as well as one or more ‘Cut repeat’ DNA binding domain(s). The cDNAs for homologues of the Drosophila melanogaster Cut homeodomain protein have been isolated from several mammalian species including human (Neufeld et al., 1992), dog (Andres et al., 1992), mouse (Valarche et al., 1993) and rat (Yoon and Chikaraishi, 1994) and were, respectively, termed CDP (CCAAT d

Cut expression in Drosophila melanogaster

In Drosophila embryos, Cut expression was observed in several tissues, and mutations affecting Cut expression or function caused developmental defects in these tissues (Table 1) (Bodmer et al., 1987, Blochlinger et al., 1988, Blochlinger et al., 1990). In the adult, cut expression was observed in the same, as well as in other, tissues (Table 1) (Blochlinger et al., 1993). Because of embryonic lethality caused by lethal cut mutations, it was not possible to determine whether cut is also required

Genetic studies in Drosophila melanogaster

A large number of viable as well as lethal cut mutations, ct, have been described in Drosophila and literature on this subject can be found as early as 1925 (reviewed in (Jack, 1985)); (Hertweck, 1931, Blanc, 1942, Braun, 1942, Bodmer et al., 1987, Jack et al., 1991, Liu et al., 1991, Jack and DeLotto, 1992, Liu and Jack, 1992). Mutations within the cut locus were divided into five groups based on the tissues affected, viability, location within the locus and complementation with other cut

Cut plays a role in cell type specification in Drosophila

Genetic studies in Drosophila melanogaster indicated that cut plays an important role in determining cell-type specificity in several tissues. Defects caused by Cut mutations appear to result from the fact that some cells have enrolled in the wrong developmental program (Bodmer et al., 1987, Blochlinger et al., 1991, Liu et al., 1991, Liu and Jack, 1992). The role of cut as a determinant of cell type specificity was most thoroughly demonstrated in the peripheral nervous system (Bodmer et al.,

The role of cut in the nervous system

While the function of cut in the central nervous system remains to be defined, the analysis of loss-of-function and gain-of-function cut mutants has revealed a role for cut in the specification of neuronal subtype in the peripheral nervous system. The formation of the peripheral nervous system in Drosophila involves several steps (reviewed in (Jan and Jan, 1994, Chan and Jan, 1999)) (Fig. 2). Sensory organs derive from one common neuronal precursor cell, which divides and differentiates to give

The role of cut in limb development: genetic interactions between cut, Notch and Wingless

Mutations that affect the expression or function of Cut, Wingless or Notch in the wing affect wing development and produce phenotypes that can be observed easily. The similitude between some of these phenotypes suggested that the products of these genes might play a role in a common process required for proper wing formation. Indeed, multiple genetic interactions have been reported between cut, and both the Notch (N) and Wingless (Wg) signaling pathways during the development of the wing

Other genetic interactions

A number of genetic interactions with cut have been reported in addition to those involved in the development of the wing. Mutations in Notch were shown to affect egg chamber formation in the ovary, however in this case a cut null mutation suppressed the notch phenotype, suggesting that the two genes have antagonistic effects in this process (Jackson and Blochlinger, 1997). In contrast, a heterozygous mutation in the Pka-C1 gene enhanced the phenotype of a cut hypomorph (Jackson and

Regulators of cut

Expression of cut in various tissues is affected by loss-of-function or ectopic expression of a number of other genes. The role of notch in the activation of cut in the wing imaginal discs has been discussed earlier. Mutations in strawberry notch (sno) (Majumdar et al., 1997), scalloped (Jack and DeLotto, 1992), mastermind (Morcillo et al., 1996, Rollins et al., 1999), Nipped-B (Rollins et al., 1999), Chip (Morcillo et al., 1997) and mod(mdg4) (Cai and Levine, 1997) were found to affect the

Biochemical activities of Cut in Drosophila

Very little information is available regarding the molecular functions of Cut proteins in Drosophila. However, Cut proteins of 215 and 230 kDa were shown to mediate ventral repression of the zen gene by binding to a sequence, AT2, containing a CDP/Cut binding site (Valentine et al., 1998). The mechanism of repression in this case is not entirely clear but may involve cooperation with the dead ringer gene product, Dri, to recruit the co-repressor Groucho.

The CAAT-displacement activity

The CCAAT-displacement activity was first described in the context of the sperm histone H2B-1 gene of the sea urchin Psammechinus miliaris (Barberis et al., 1987). This gene is expressed in testis but not in the rest of the embryo. Its lack of expression in tissues other than testis was found to correlate with the presence of a factor that was capable of displacing the ubiquitous CCAAT binding factor (CBF) from the promoter. Analysis of the γ-globin gene promoter, that differs from the other

CDP/Cut DNA binding activity in mammals generally correlates with cellular proliferation

The bulk of the results in mammals suggested that CDP/Cut expression or activity might be restricted to proliferating cells. Expression of the mouse CDP/Cut protein, Cux-1, in the kidney was found to be inversely related to the degree of cellular differentiation (Heuvel et al., 1996). CDP/Cut binding activity, initially characterized as the CCAAT displacement activity (Skalnik et al., 1991, Lievens et al., 1995) or histone nuclear factor D (HiNF-D) (van Wijnen et al., 1989), was found to

Mechanisms of repression by CDP/Cut

CDP/Cut proteins were found generally to function as transcriptional repressors (Superti-Furga et al., 1989, Skalnik et al., 1991, Andres et al., 1992, Valarche et al., 1993, Dufort and Nepveu, 1994, Lievens et al., 1995, Mailly et al., 1996, Li et al., 1999, van Gurp et al., 1999). Repression by CDP/Cut involves at least two mechanisms. In most cases, CDP/Cut appears to compete with some activators for the occupancy of a binding site. It is this property that bestowed upon this protein the

CDP/Cut may also function as an activator

The role of CDP/Cut in proliferating cells may not be limited to that of a transcriptional repressor, as certain results suggest that CDP/Cut may also be able to participate in gene activation. A number of groups have identified binding sites for CDP/Cut in the regulatory sequences of genes encoding for histones and the thymidine kinase (Barberis et al., 1987, el-Hodiri and Perry, 1995, van Wijnen et al., 1996, Kim et al., 1997). The peak of expression of these genes coincides with or closely

Matrix attachment regions (MAR)

A number of reports have described the interaction of CDP/Cut with matrix attachment regions (MARs) that are believed to play an important role in gene regulation. These include the MARs upstream of the CD8α (Banan et al., 1997) and T cell receptor β genes (Chattopadhyay et al., 1998), the MARs flanking the immunoglobulin heavy chain intronic enhancer (Wang et al., 1999), the negative regulatory elements within the mouse mammary tumor virus (MMTV) long terminal repeat (LTR) (Liu et al., 1997),

DNA binding domains of CDP/Cut

The high degree of conservation of Cut repeats suggested that they might have an important biochemical function (Blochlinger et al., 1988, Neufeld et al., 1992). Indeed, Cut repeats were found to function as specific DNA binding domains (Andres et al., 1994, Aufiero et al., 1994, Harada et al., 1994, Harada et al., 1995). Cut proteins therefore are unique in that they contain four DNA binding domains: the Cut homeodomain and the three Cut repeats. How exactly the CDP/Cut protein interacts with

CDP/Cut DNA binding is inhibited by post-translational modifications of either cut repeats or the cut homeodomain

Comparison with the sequence of Drosophila Cut revealed that Cut repeats contain evolutionarily conserved consensus phosphorylation sites for protein kinase C (PKC) and casein kinase II (CKII). PKC (Coqueret et al., 1996) and CKII (Coqueret et al., 1998b) were shown to phosphorylate Cut repeats, in vitro and in vivo. Phosphorylation of Cut repeats caused an inhibition of DNA binding and, consequently, the transcriptional repression activity of CDP/Cut. In fibroblastic cells, CDP/Cut DNA binding

The existence of Cux-1 and Cux-2 genes in vertebrates suggests some degree of redundancy and specialization

A second Cux gene, called Cux-2, was identified in mouse and chicken (Quaggin et al., 1996, Tavares et al., 2000). An almost identical human cDNA sequence has been isolated from a human brain library, suggesting that a second, neuronal specific, CDP/Cut gene exists in all vertebrates (NCBI, accession number AB006631). In contrast to mouse Cux-1 which is expressed in most tissues (including the nervous system), Cux-2 was found to be expressed primarily in nervous tissues (Quaggin et al., 1996).

Genetic studies in mouse

An early attempt to generate a knock-out mouse resulted in the generation of a mouse expressing a Cux protein with an internal deletion of 246 amino acids encompassing Cut repeat 1. DeltaCR1 homozygous mutant mice displayed a mild phenotype characterized by curly vibrissae, wavy hair, and a high degree of pup loss resulting probably from feeding problems (Tufarelli et al., 1998). More recently, the group of Dr Ellis J. Neufeld has generated another knock-out, designated DeltaHD, in which the

CUTL1 is located in a chromosomal region that is frequently rearranged in cancers

The CUTL1 gene was mapped to chromosome 7, band 7q22 (Scherer et al., 1993, Lemieux et al., 1994). Cytogenetic analyses revealed that rearrangements or deletions of 7q22 occur frequently in uterine leiomyomas (reviewed in (Ozisik et al., 1993), acute myeloid leukemia (Fenaux et al., 1989, Swansbury et al., 1994) and myelodysplastic syndrome (Yunis et al., 1988, Heim, 1992). The fact that such a high proportion of some cancers present a cytogenetically detectable deletion of 7q22 suggested that

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ReviewRole of the multifunctional CDP/Cut/Cux homeodomain transcription factor in regulating differentiation, cell growth and development

Abstract

Section snippets

CDP/Cut is conserved among metazoans

Cut expression in Drosophila melanogaster

Genetic studies in Drosophila melanogaster

Cut plays a role in cell type specification in Drosophila

The role of cut in the nervous system

The role of cut in limb development: genetic interactions between cut, Notch and Wingless

Other genetic interactions

Regulators of cut

Biochemical activities of Cut in Drosophila

The CAAT-displacement activity

CDP/Cut DNA binding activity in mammals generally correlates with cellular proliferation

Mechanisms of repression by CDP/Cut

CDP/Cut may also function as an activator

Matrix attachment regions (MAR)

DNA binding domains of CDP/Cut

CDP/Cut DNA binding is inhibited by post-translational modifications of either cut repeats or the cut homeodomain

The existence of Cux-1 and Cux-2 genes in vertebrates suggests some degree of redundancy and specialization

Genetic studies in mouse

CUTL1 is located in a chromosomal region that is frequently rearranged in cancers

J. Biol. Chem.

Cell

Curr. Opin. Genet. Dev.

Cell

Curr. Opin. Neurobiol.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Trends Genet.

J. Biol. Chem.

Leukemia Res.

Biochim. Biophys. Acta – Gene Struct. Express.

Curr. Opin. Cell. Biol.

Cell

J. Biol. Chem.

Mech. Dev.

Curr. Biol.

Blood

Gene

Blood

Genomics

J. Biol. Chem.

J. Biol. Chem.

Dev. Biol.

Dev. Biol.

J. Biol. Chem.

Neuron

J. Biol. Chem.

Cancer Genet. Cytogenet.

J. Biol. Chem.

Genomics

J. Biol. Chem.

Biochim. Biophys. Acta

Cancer Genet. Cytogenet.

CCAAT displacement protein binds to and negatively regulates human papillomavirus type 6 E6, E7, and E1 promoters

J. Virol.

A new bipartite DNA-binding domain: cooperative interaction between the cut repeat and homeo domain of the cut homeo proteins

Genes Dev.

Clox, a mammalian homeobox gene related to Drosophila cut, encodes DNA-binding regulatory proteins differentially expressed during development

Development

Notch signaling: cell fate control and signal integration in development

Science

Sequence-specific DNA binding of individual Cut repeats of the human CCAAT displacement/Cut homeodomain protein

Proc. Natl. Acad. Sci. USA

HiNF-D (CDP-cut/CDC2/cyclin A/pRB-complex) influences the timing of IRF- 2-dependent cell cycle activation of human histone H4 gene transcription at the G1/S phase transition

J. Cell. Physiol.

The integrated activities of Irf-2 (Hinf-M), Cdp/Cut (Hinf-D) and H4tf-2 (Hinf-P) regulate transcription of a cell cycle controlled human histone H4 gene – mechanistic differences between distinct H4 genes

Mol. Biol. Rep.

The production of wing scalloping in Drosophila melanogaster

Univ. Calif. Publ. Zool.

Primary structure and expression of a product from cut, a locus involved in specifying sensory organ identity in Drosophila

Nature

Patterns of expression of cut, a protein required for external sensory organ development in wild-type and cut mutant Drosophila embryos

Genes Dev.

Transformation of sensory organ identity by ectopic expression of Cut in Drosophila

Genes Dev.

Postembryonic patterns of expression of cut, a locus regulating sensory organ identity in Drosophila

Review
Role of the multifunctional CDP/Cut/Cux homeodomain transcription factor in regulating differentiation, cell growth and development