Mutations that affect the ability of the vnd/NK-2 homeoprotein to regulate gene expression: Transgenic alterations and tertiary structure

Koizumi, Keita; Lintas, Carla; Nirenberg, Marshall W.; Maeng, Jin-Soo; Ju, Jeong-Ho; Mack, James W.; Gruschus, James M.; Odenwald, Ward F.; Ferretti, James A.

doi:10.1073/pnas.0438043100

Cited by 25 publications

(30 citation statements)

References 39 publications

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“…For instance, Pax5 and Bap interact with Gro/TLE through sequences besides their octapeptide/eh1 domains, and AML1 lacking its WRPY motif retains some Gro-binding activity (11,21,40). Importantly, the occurrence of multiple Gro-binding sites within repressors might explain why mutations in some eh1 domains lead to surprisingly mild phenotypes (12,16,39). It is conceivable that synergistic interactions between multiple weak binding domains within repressors and in Gro/TLE provide composite interfaces that allow for high-affinity protein interactions and hence for robust repressor capability in vivo.…”

Section: Recruitment Of Gro By Repressors Containing Eh1-like Motifsmentioning

confidence: 99%

“…In this report, we have shown that eh1 motifs are prevalent among members of various transcription factor families and have further demonstrated that one of these, specifically, the ZnF protein Odd, requires its eh1 Gro-binding domain for blocking gene expression in vivo. While our work was in progress, three additional eh1-containing proteins, shown in this paper to complex with Gro in vitro, were found to behave as repressors in vivo, i.e., Slp (1, 2), BarH1 (41), and Vnd (16,39). These reports independently support our conclusions and, along with the results presented here, suggest that a significant number of uncharacterized transcriptional regulators, containing eh1-related motifs, function as Gro-dependent repressors.…”

Section: Recruitment Of Gro By Repressors Containing Eh1-like Motifsmentioning

confidence: 99%

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An eh1-Like Motif in Odd-skipped Mediates Recruitment of Groucho and Repression In Vivo

Goldstein

Cook

Dinur

et al. 2005

Molecular and Cellular Biology

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Drosophila Groucho, like its vertebrate Transducin-like Enhancer-of-split homologues, is a corepressor that silences gene expression in numerous developmental settings. Groucho itself does not bind DNA but is recruited to target promoters by associating with a large number of DNA-binding negative transcriptional regulators. These repressors tether Groucho via short conserved polypeptide sequences, of which two have been defined. First, WRPW and related tetrapeptide motifs have been well characterized in several repressors. Second, a motif termed Engrailed homology 1 (eh1) has been found predominantly in homeodomain-containing transcription factors. Here we describe a yeast two-hybrid screen that uncovered physical interactions between Groucho and transcription factors, containing eh1 motifs, with different types of DNA-binding domains. We show that one of these, the zinc finger protein Odd-skipped, requires its eh1-like sequence for repressing specific target genes in segmentation. Comparison between diverse eh1 motifs reveals a bias for the phosphoacceptor amino acids serine and threonine at a fixed position, and a mutational analysis of Oddskipped indicates that these residues are critical for efficient interactions with Groucho and for repression in vivo. Our data suggest that phosphorylation of these phosphomeric residues, if it occurs, will down-regulate Groucho binding and therefore repression, providing a mechanism for posttranslational control of Grouchomediated repression.Negative transcriptional regulation is a strategy that has been commonly selected in evolution for setting up and maintaining gene expression patterns. A striking case in point is the process of segmentation in the early Drosophila embryo. This developmental system is regulated almost exclusively by transcription factors, many of which are repressors that silence the expression of their targets (35; reviewed in reference 57). Mutations in genes encoding these transcriptional repressors lead to the loss of repressor activity that normally restricts the expression domains of downstream genes, causing disruptions in the metameric subdivision of the fly embryo.One key principle to emerge from studies on Drosophila segmentation and on developmental processes in other model organisms is the fact that DNA-binding repressors in general do not operate on their own. Instead, they complex with nuclear coregulators, called corepressors, tethering them to promoters whose expression is subsequently blocked (43). Groucho (Gro), one such ubiquitously expressed corepressor that is highly conserved throughout evolution from worms to humans, has been shown to interact with and to potentiate the repressor function of a vast number of transcription factors, including many of those acting in segmentation (8). It is not fully understood how Gro and its Transducin-like Enhancer-of-split (TLE) mammalian homologues elicit transcriptional repression, although given that these corepressors associate with histones and bind histone deacetylases, they are likely to ...

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Section: Recruitment Of Gro By Repressors Containing Eh1-like Motifsmentioning

confidence: 99%

Section: Recruitment Of Gro By Repressors Containing Eh1-like Motifsmentioning

confidence: 99%

An eh1-Like Motif in Odd-skipped Mediates Recruitment of Groucho and Repression In Vivo

Goldstein

Cook

Dinur

et al. 2005

Molecular and Cellular Biology

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“…In principle, ind can be activated in both regions by the combination of low levels of the Dorsal gradient and EGF signaling, but expression is kept off in the ventral neurogenic ectoderm by Vnd, a sequencespecific transcriptional repressor that is activated by intermediate levels of Dorsal ( Fig. 1) (23)(24)(25). As a result of this repression, ind expression is restricted to lateral regions of the neurogenic ectoderm.…”

Section: Cell-cell Interactions Produce Additional Dorsal Gradient Rementioning

confidence: 99%

How the Dorsal gradient works: Insights from postgenome technologies

Hong

Hendrix

Papatsenko

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

123

128

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Gradients of extracellular signaling molecules and transcription factors are used in a variety of developmental processes, including the patterning of the Drosophila embryo, the establishment of diverse neuronal cell types in the vertebrate neural tube, and the anterior-posterior patterning of vertebrate limbs. Here, we discuss how a gradient of the maternal transcription factor Dorsal produces complex patterns of gene expression across the dorsal-ventral (DV) axis of the early Drosophila embryo. The identification of 60 -70 Dorsal target genes, along with the characterization of Ϸ35 associated regulatory DNAs, suggests that there are at least six different regulatory codes driving diverse DV expression profiles.Drosophila ͉ regulatory code ͉ embryo ͉ morphogen T he dorsal-ventral (DV) patterning of the early Drosophila embryo is controlled by a sequence-specific transcription factor, Dorsal, which is related to mammalian NF-B (1-3). Differential activation of the Toll receptor leads to the formation of a broad Dorsal nuclear gradient, with peak levels present in ventral regions and progressively lower levels in lateral and dorsal regions (Fig.

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“…Previously, it has been reported that the amino acid residues at the positions 3 and 5 (Vnd 547 and 549, respectively) of the N-terminal arm as well as residues at the positions 47, 50, and 54 (Vnd 591, 595, and 598, respectively) of the recognition helix in HD directly contact with the bases of the DNA (5,9,16). In addition to the residues directly involved in base-specific interactions, amino acid side chains at the positions that do not contact any DNA bases are believed to affect the interaction of the HD with its target DNA, and thus may be important functionally.…”

Section: Kinetic Analysis Of Alanine Substitutions Within Either Homementioning

confidence: 99%

Kinetic analysis of Drosophila Vnd protein containing homeodomain with its target sequence

Yoo

2010

BMB Reports

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Homeodomain (HD) is a highly conserved DNA-binding domain composed of helix-turn-helix motif. Drosophila Vnd (Ventral nervous system defective) containing HD acts as a regulator to either enhance or suppress gene expression upon binding to its target sequence. In this study, kinetic analysis of Vnd binding to DNA was performed. The result demonstrates that DNA-binding affinity of the recombinant protein containing HD and NK2-specific domain (NK2-SD) was higher than that of the full-length Vnd. To access whether phosphorylation sites within HD and NK2-SD affect the interaction of the protein with the target sequence, alanine substitutions were introduced. The result shows that S631A mutation within NK2-SD does not contribute significantly to the DNA-binding affinity. However, S571A and T600A mutations within HD showed lower affinity for DNA binding. In addition, DNA-binding analysis using embryonic nuclear protein also demonstrates that Vnd interacts with other nuclear proteins, suggesting the existence of Vnd as a complex. [BMB reports 2010; 43(6): 407-412]

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Mutations that affect the ability of the vnd/NK-2 homeoprotein to regulate gene expression: Transgenic alterations and tertiary structure

Cited by 25 publications

References 39 publications

An eh1-Like Motif in Odd-skipped Mediates Recruitment of Groucho and Repression In Vivo

An eh1-Like Motif in Odd-skipped Mediates Recruitment of Groucho and Repression In Vivo

How the Dorsal gradient works: Insights from postgenome technologies

Kinetic analysis of Drosophila Vnd protein containing homeodomain with its target sequence

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