Acquisition of NFKB1-selective DNA binding by substitution of four amino acid residues from NFKB1 into RelA.

Coleman, T A; Kunsch, C; Maher, M; Ruben, S M; Rosen, C A

doi:10.1128/mcb.13.7.3850

Cited by 20 publications

(15 citation statements)

References 44 publications

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“…Unlike most transcription factors which use alpha helices to bind DNA, the Rel/NF-kB proteins use ten¯exible loops extending from the secondary structure of these immunoglobulin folds to mediate DNA contacts. These structures support numerous biochemical results that indicate the N' region of the RH domain is important for DNA binding (Coleman et al, 1993;Toledano et al, 1993).…”

Section: Structure Of Nf-kb/dna Complexessupporting

confidence: 84%

“…HrdlickovaÂ et al (1995) have mapped the altered DNA-binding speci®city to the mutations (from c-Rel to v-Rel) at Met20Thr, Asp82Gly, and Arg92Glu. Previous experimental evidence has shown that Met20Thr is one of four residues which are responsible for di erent DNAbinding speci®cities between p50 and RelA (Coleman et al, 1993). Furthermore, Nehyba et al (1997) have also shown that Met20Thr in v-Rel alters DNA-binding speci®cities and have provided additional evidence that the Arg250Gly and Glu269Ala mutations in v-Rel cause promiscuous kB site binding.…”

Section: Protein Sequence Variationmentioning

confidence: 95%

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Regulation of DNA binding by Rel/NF-κB transcription factors: structural views

1999

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Rel/NF-kB transcription factors form homo-and heterodimers with di erent DNA binding site speci®cities and DNA binding a nities. Several intracellular pathways evoked by a wide range of biological factors and environmental conditions can lead to the activation of Rel/NF-kB dimers by signaling degradation of the inhibitory IkB protein. In the nucleus Rel/NF-kB dimers modulate the expression of a variety of genes including those encoding cytokines, growth factors, acute phase response proteins, immunoreceptors, other transcription factors, cell adhesion molecules, viral proteins and regulators of apoptosis. The primary focus of this review is on structural and functional aspects of Rel/NFkB:DNA complexes and their formation. The salient features of the Rel/NF-kB dimer:DNA structure are described, as are modes of transcriptional regulation by phosphorylation, altered DNA binding properties, varying protein conformations, and interactions with IkB proteins.

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Section: Structure Of Nf-kb/dna Complexessupporting

confidence: 84%

Section: Protein Sequence Variationmentioning

confidence: 95%

Regulation of DNA binding by Rel/NF-κB transcription factors: structural views

1999

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“…Met-20?Thr is located in the major DNA recognition loop in the RxxRxRxxC binding motif of v-Rel (Kumar et al, 1992). This mutation resides in one of the four amino acid sequence positions demonstrated to be important in de®ning the di erence in the DNAbinding speci®city of human RelA and p50 (NF-kB1) (Coleman et al, 1993;Toledano et al, 1993). Interestingly, it is only 10 amino acids from the mutation Ala-31?Ser, which was identi®ed in a vRel mutant with higher transformation e ciency for Bcells (Romero and Humphries, 1995).…”

Section: N-rh Mutationsmentioning

confidence: 99%

Differences in κB DNA-binding properties of v-Rel and c-Rel are the result of oncogenic mutations in three distinct functional regions of the Rel protein

1997

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The oncogene v-rel of Reticuloendotheliosis virus, strain T, is derived from an avian c-rel proto-oncogene. c-rel encodes a member of the Rel/NF-kB family of transcription factors. The highly oncogenic v-Rel di ers from c-Rel which has low transforming potential by the acquisition of numerous mutations. In this manuscript, we demonstrate that the oncogenic mutations in v-Rel directly alter the ability of this protein to bind to DNA. Electrophoretic mobility shift analysis with Rel proteins synthesized in vitro as well as isolated from nuclei of Rel expressing cells showed that three mutation clusters, present in the N-terminus, the center and the C-terminus of v-Rel, altered three di erent aspects of DNA binding. In contrast, the oncogenic C-terminal deletion of 118 amino acids present in v-Rel had almost no in¯uence on its DNA binding. The N-terminal mutation cluster altered the kB DNA-binding speci®city of the v-Rel oncoprotein relative to c-Rel. The mutation Met-20?Thr was found to be principally responsible for this alteration. The second mutation cluster was responsible for increased binding of v-Rel to all the kB sites examined presumably because it stabilized v-Rel homodimers. This alteration in DNA binding was mapped to the group of two mutations within the cluster. In contrast, the third mutation cluster in the C-terminus of v-Rel destabilized the binding of v-Rel to all of the kB sites examined. This is the ®rst indication that regions outside the Rel Homology Region can participate in the control of binding of the c-Rel protein to DNA. The three mutation clusters examined contributed to the tumorigenic potential of v-Rel with the relative strength decreasing with their position from the N-terminus to the C-terminus. These results suggest that the oncogenic mutations in v-Rel cooperate and enable v-Rel to form nuclear complexes with aberrant DNA-binding properties that may directly alter gene expression and DNA replication resulting in the transformation of the cell.

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“…A shorter alternatively spliced transcript of NF-KB2 has also been identified; it gives rise to a 49-kDa protein, NF-KB2(p49), identical to the amino-terminal Rel domain of NF-KB2(plOO), and thus does not require processing to generate the DNA-binding form (62) but otherwise has activities identical to those of NF-KB2(p52) (54). The DNA-binding and dimerization functions of NF-KB subunits have been shown to be encoded by the amino-and carboxyterminal regions, respectively, of the Rel-conserved domain (11,13,35,42,73). Deletion of the amino-terminal region of NF-KB1(pSO) abolishes DNA binding while leaving dimerization function intact.…”

mentioning

confidence: 99%

“…Deletion of the amino-terminal region of NF-KB1(pSO) abolishes DNA binding while leaving dimerization function intact. Furthermore, mutations within the first 43 amino acids of RelA have been shown to alter DNA-binding specificity (13), although precise designation of the residues which determine DNA binding and dimerization will require structural analysis of the Rel homology domain.…”

mentioning

confidence: 99%

An interaction between the DNA-binding domains of RelA(p65) and Sp1 mediates human immunodeficiency virus gene activation.

Perkins¹,

Agranoff²,

Pascal³

et al. 1994

Mol. Cell. Biol.

225

157

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Induction of human immunodeficiency virus type 1 (HIV-1) gene expression in stimulated T cells has been attributed to the activation of the transcription factor NF-KB. The twice-repeated KB sites within the HIV-1 long terminal repeat are in close proximity to three binding sites for Spl. We have previously shown that a cooperative interaction of NF-KB with Spl is required for the efficient stimulation of HIV-1 transcription. In this report, we define the domains of each protein responsible for this effect. Although the transactivation domains seemed likely to mediate this interaction, we find, surprisingly, that this interaction occurs through the putative DNA-binding domains of both proteins. Spl specifically interacted with the amino-terminal region of ReIA(p65). Similarly, RelA bound directly to the zinc finger region of Spl. This interaction was specific and resulted in cooperative DNA binding to the icB and Spl sites in the HIV-1 long terminal repeat. Furthermore, the amino-terminal region of RelA did not associate with several other transcription factors, including MyoD, E12, or Koxl5, another zinc finger protein. These findings suggest that the juxtaposition of DNA-binding sites promotes a specific protein interaction between the DNA-binding regions of these transcription factors. This interaction is required for HIV transcriptional activation and may provide a mechanism to allow for selective activation of KB-regulated genes.

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Acquisition of NFKB1-selective DNA binding by substitution of four amino acid residues from NFKB1 into RelA.

Cited by 20 publications

References 44 publications

Regulation of DNA binding by Rel/NF-κB transcription factors: structural views

Regulation of DNA binding by Rel/NF-κB transcription factors: structural views

Differences in κB DNA-binding properties of v-Rel and c-Rel are the result of oncogenic mutations in three distinct functional regions of the Rel protein

An interaction between the DNA-binding domains of RelA(p65) and Sp1 mediates human immunodeficiency virus gene activation.

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