Biochemical Analysis of the Kruppel-associated Box (KRAB) Transcriptional Repression Domain

Peng, Hongzhuang; Begg, Gillian E.; Harper, Sandra L.; Friedman, Josh R.; Speicher, David W.; Rauscher, Frank J.

doi:10.1074/jbc.m001499200

Cited by 100 publications

(94 citation statements)

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“…It is located at the N terminus when it is part of a zinc finger-containing transcription factor. The primary amino acid sequence of the domain does not resemble any of the other zinc finger-associated domains, such as the Kruppelassociated box (KRAB) or the poxvirus and zinc finger (POZ) domain, which is also known as the BTB (Broad-complex, Tramtrack, and Bric-a-brac) domain (7)(8)(9). Members of the SCAN domain family are broadly expressed and appear to function as either activators or repressors of transcription.…”

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confidence: 99%

The SCAN Domain of ZNF174 Is a Dimer

Stone¹,

Maki²,

Blacklow

et al. 2002

Journal of Biological Chemistry

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The SCAN domain is a conserved region of 84 residues found predominantly in zinc finger DNA-binding proteins in vertebrates. The SCAN domain appears to control the association of SCAN domain containing proteins into noncovalent complexes and may be the primary mechanism underlying partner choice in the oligomerization of these transcription factors. Here we have overexpressed, purified, and characterized the isolated SCAN domain (amino acids 37-132) from ZNF174. Both size exclusion chromatography and equilibrium sedimentation analysis demonstrate that the ZNF174 SCAN domain forms a homodimer. Circular dichroism shows that the isolated SCAN domain dimer has ϳ42% ␣-helix. Thermal denaturation experiments indicate that the SCAN domain undergoes a single reversible unfolding transition with a T m of over 70°C. The midpoint of the equilibrium unfolding transition increases with increasing protein concentration, consistent with a two-state unfolding transition in which folded dimer is in equilibrium with unfolded monomer. These findings demonstrate that the isolated SCAN domain forms a stable dimer and support a model in which the SCAN domain is capable of mediating the selective dimerization of a large family of vertebrate-specific, zinc finger-containing transcription factors.Transcription factors are composed of modular elements that include a DNA-binding domain and one or more separable effector domains that activate or repress transcription. Other modules within these factors regulate subcellular localization and gene expression by mediating selective association of the transcription factors with each other, or with other cellular components. Identification of these domains often provides a conceptual framework for understanding the function of the transcription factor.The SCAN 1 domain is a highly conserved vertebrate-specific protein domain found in ϳ60 genes in the human genome (1, 2). Only a handful of these gene products have been characterized thus far, and they appear to control a wide range of biological processes including development, cell differentiation, and lipid metabolism (reviewed in Refs. 3 and 4). The name for the SCAN domain was derived from the first letters of the names of four proteins initially found to contain this domain (SRE-ZBP, CTfin51, AW-1 (ZNF174), and Number 18 cDNA or ZNF197) (5). Alternatively, this domain has been referred to as LeR for leucine-rich domain (6). The SCAN domain consists of an 84-residue, leucine-rich region and is predicted to contain a high degree of ␣-helix. It is located at the N terminus when it is part of a zinc finger-containing transcription factor. The primary amino acid sequence of the domain does not resemble any of the other zinc finger-associated domains, such as the Kruppelassociated box (KRAB) or the poxvirus and zinc finger (POZ) domain, which is also known as the BTB (Broad-complex, Tramtrack, and Bric-a-brac) domain (7-9). Members of the SCAN domain family are broadly expressed and appear to function as either activators or repressors of transcri...

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confidence: 99%

The SCAN Domain of ZNF174 Is a Dimer

Stone¹,

Maki²,

Blacklow

et al. 2002

Journal of Biological Chemistry

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“…There exist a number of publications describing the correlation between the expression of ZNF280B and the growth of various types of cancer (5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22). For example, Gunn et al (22) reported that ZNF280B acts as a tumor suppressor in chronic lymphocytic leukemia.…”

Section: Discussionmentioning

confidence: 99%

“…By interacting with DNA and proteins, ZNF proteins can provide links between these molecule types (4). An increasing number of studies has reported that ZNF proteins serve critical roles in human cancers (5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15). For example, ZNF703 acts as an oncogene in invasive gastric carcinoma, and its expression levels are correlated with gastric carcinoma progression (16).…”

Section: Introductionmentioning

confidence: 99%

ZNF280B promotes the growth of gastric cancer inï¿½vitro and inï¿½vivo

et al. 2018

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Abstract. Zinc finger protein 280B (ZNF280B) mediates pro-survival and pro-growth functions in prostate cancer. However, in gastric cancer, its clinical significance remains poorly characterized. In the present study, the expression levels of ZNF280B in 60 patients with gastric cancer were examined using immunohistochemistry. The association between ZNF280B expression and clinicopathological features was assessed. Positive ZNF280B staining was demonstrated for 38 (63.3%) samples out of 60 gastric cancer cases in immunohistochemical analysis. ZNF280B expression was significantly associated with tumor size (P= 0.017) and TNM stage (P= 0.001). Furthermore, the proliferation index in the positive ZNF280B expression group was significantly higher (38.8±6.2) compared with that of the negative ZNF280B expression group (16.9±8.9; P<0.01). These results suggest that ZNF280B expression may be associated with the proliferation of gastric cancer cells. The role of ZNF280B in the growth of gastric cancer cells (MGC-803) was also investigated in vitro and in vivo by enhancing the expression of ZNF280B. A colony formation assay indicated that the number of colonies in the MGC-803 cells with enhanced ZNF280B (146±5.8) was significantly higher than that of the MGC-803 control group (97±5.1) and the negative control group (101±6.5; P<0.05). An MTT assay demonstrated that ZNF280B significantly promoted the proliferation of MGC-803 cells at days 3 and 4 (P<0.05). It was observed that the overexpression of ZNF280B may promote the growth of gastric cancer in vivo in xenograft studies. These findings indicate that ZNF280B may be a novel therapeutic target for gastric cancer.

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“…Repressor functions are often mediated through an interaction of the repressor domain with nuclear co-repressors. For example, the repression by the Krü ppel-associated box is mediated through interaction with the co-repressor TIF1 (4,5,65). As mentioned above, the speckled-like distribution of Glis1 in the nucleus suggests association with a larger protein complex and possibly a co-repressor complex.…”

Section: Figmentioning

confidence: 99%

“…subfamilies based on the number of zinc finger motifs, sequence homology between the zinc-fingers, and the presence of specific repressor and activation domains (3)(4)(5)(6).…”

mentioning

confidence: 99%

Identification of Glis1, a Novel Gli-related, Krüppel-like Zinc Finger Protein Containing Transactivation and Repressor Functions

Kim

Lewandoski

Perantoni

et al. 2002

Journal of Biological Chemistry

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In this study, we describe the identification and characterization of a novel Krü ppel-like protein named Glisimilar 1 (Glis1). The Glis1 gene encodes an 84.3-kDa proline-rich protein. Its five tandem zinc finger motifs exhibit highest homology with those of members of the Gli and Zic subfamilies of Krü ppel-like proteins. Glis1 was mapped to mouse chromosome 4C6. Northern blot analysis showed that expression of the 3.3-kb Glis1 mRNA is most abundant in placenta and adult kidney and expressed at lower levels in testis. Whole mount in situ hybridization on mouse embryos demonstrated that Glis1 is expressed in a temporal and spatial manner during development; expression was most prominent in several defined structures of mesodermal lineage, including craniofacial regions, branchial arches, somites, vibrissal and hair follicles, limb buds, and myotomes. Confocal microscopic analysis showed that Glis1 is localized to the nucleus. The zinc finger region plays an important role in the nuclear localization of Glis1. Electrophoretic mobility shift assays demonstrated that Glis1 is able to bind oligonucleotides containing the Glibinding site consensus sequence GACCACCCAC. Although monohybrid analysis showed that in several cell types Glis1 was unable to induce transcription of a reporter, deletion mutant analysis revealed the presence of a strong activation function at the carboxyl terminus of Glis1. The activation through this activation function was totally suppressed by a repressor domain at its amino terminus. Constitutively active Ca 2؉ -dependent calmodulin kinase IV enhanced Glis1-mediated transcriptional activation about 4-fold and may be mediated by phosphorylation/activation of a co-activator. Our results suggest that Glis1 may play a critical role in the control of gene expression during specific stages of embryonic development.

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Biochemical Analysis of the Kruppel-associated Box (KRAB) Transcriptional Repression Domain

Cited by 100 publications

References 50 publications

The SCAN Domain of ZNF174 Is a Dimer

The SCAN Domain of ZNF174 Is a Dimer

ZNF280B promotes the growth of gastric cancer inï¿½vitro and inï¿½vivo

Identification of Glis1, a Novel Gli-related, Krüppel-like Zinc Finger Protein Containing Transactivation and Repressor Functions

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