Structure of serum response factor core bound to DNA

Pellegrini, Luca; Tan, Song; Richmond, Timothy J.

doi:10.1038/376490a0

Cited by 320 publications

(418 citation statements)

References 47 publications

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“…Blake (41) suggested that evidence for intron-facilitated evolution of a gene might be found by comparing the borders of functional protein domains with the placement of introns. The recent elucidation of the x-ray crystal structure of the SRF MADS box demonstrated a novel DNA-binding motif, a coiled-coil, and a stratified structural subdomain involved in dimerization (2). We showed here ( Fig.…”

Section: Fig 9 Srf Promoter Activity Elevated During Myogenesis Is mentioning

confidence: 52%

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Organization and Myogenic Restricted Expression of the Murine Serum Response Factor Gene

Belaguli

Schildmeyer

Schwartz

1997

Journal of Biological Chemistry

134

121

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Serum response factor (SRF), a member of an ancient family of DNA-binding proteins, is generally assumed to be a ubiquitous transcription factor involved in regulating growth factor-responsive genes. However, avian SRF was recently shown (Croissant, J. D., Kim, J.-H., Eichele, G., Goering, L., Lough, J., Prywes, R., and Schwartz, R. J. (1996) Dev. Biol. 177, 250 -264) to be preferentially expressed in myogenic lineages and is required for regulating post-replicative muscle gene expression. Given the central importance of SRF for the muscle tissue-restricted expression of the striated ␣-actin gene family, we wanted to determine how SRF might contribute to this muscle-restricted expression. Here we have characterized the murine SRF genomic locus, which has seven exons interrupted by six introns, with the entire locus spanning 11 kilobases. Murine SRF transcripts were processed to two 3 -untranslated region polyadenylation signals, yielding 4.5-and 2.5-kilobase mRNA species. Murine SRF mRNA levels were the highest in adult skeletal and cardiac muscle, but barely detected in liver, lung, and spleen tissues. During early mouse development, in situ hybridization analysis revealed enrichment of SRF transcripts in the myotomal portion of somites, the myocardium of the heart, and the smooth muscle media of vessels of mouse embryos. Likewise, murine SRF promoter activity was tissue-restricted, being 80-fold greater in primary skeletal myoblasts than in liver-derived HepG2 cells. In addition, SRF promoter activity increased 6-fold when myoblasts withdrew from the cell cycle and fused into differentiated myotubes. A 310-base pair promoter fragment depended upon multiple intact serum response elements in combination with Sp1 sites for maximal myogenic restricted activity. Furthermore, cotransfected SRF expression vector stimulated SRF promoter transcription, whereas dominant-negative SRF mutants blocked SRF promoter activity, demonstrating a positive role for an SRF-dependent autoregulatory loop. Earlier studies (8 -10) have demonstrated that SRF-binding sites, termed serum response elements (SRE; GG(A/T) 6 CC), play a primary role in regulating early response genes such as c-fos and egr-1 (reviewed in Ref. 11). Growth factor signaling through the c-fos SRE appeared to be mediated through the formation of ternary complexes with an accessory factor, p62 TCF , and through protein-DNA interactions with a purinerich sequence at the 5Ј-end of the c-fos SRE (12, 13). The ETS domain proteins Elk-1 (14) and SAP-1 (15) possess biochemical activities that are characteristic of p62 TCF (16) and interact with the C-terminal portion of the MADS box (17). Furthermore, another MADS box accessory factor, a paired-like homeodomain protein, Phox1 (18), has been shown to facilitate the DNA binding activity of SRF on the c-fos SRE. In addition, Phox1 and Elk-1 potentiate the ability of SRF to transcriptionally activate the c-fos promoter in response to growth-mediated events (18).SRF plays an obligatory role in regulating post-replicative muscl...

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Section: Fig 9 Srf Promoter Activity Elevated During Myogenesis Is mentioning

confidence: 52%

“…The structure of the MADS box domain, recently elucidated by Pellegrini et al (2), was assembled before the divergence of plants and animals. The identical MADS box structures were present in yeast transcription factors MCM1 and ARG80, a large number of homeotic like plant proteins, and invertebrate and vertebrate SRFs (reviewed in Ref.…”

mentioning

confidence: 99%

Organization and Myogenic Restricted Expression of the Murine Serum Response Factor Gene

Belaguli

Schildmeyer

Schwartz

1997

Journal of Biological Chemistry

134

121

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“…The N-terminal DNA-contacting half of the MADS domain forms an amphipathic a-helix that contacts DNA and contributes to the strength of dimerization. The C-terminal half of MADS domain forms a series of anti-parallel b-sheets that function as a dimerization interface (Hassler and Richmond, 2001;Pellegrini et al, 1995;Santelli and Richmond, 2000;Tan and Richmond, 1998). We did not address critical amino acids in the MADS domain in the reverse yeast two-hybrid screens due to the fact that MADS domain-containing versions of AP3 and PI do not interact strongly in yeast two-hybrid assays.…”

Section: Discussionmentioning

confidence: 99%

“…The 56-amino acid MADS domain possesses both DNA-binding and dimerization functions as demonstrated by X-ray structures of the MADS domains of the mammalian proteins SRF and MEF2 and the yeast protein MCM1. The amino terminal end of the MADS domain contacts DNA while the C-terminal end contains two anti-parallel b sheets that function as a dimerization interface (Figure 1) (Hassler and Richmond, 2001;Pellegrini et al, 1995;Santelli and Richmond, 2000;Tan and Richmond, 1998). The 80-amino acid K domain contains several (abcdefg) n heptad repeats in which a and d positions are occupied by hydrophobic amino acids suggesting that K domain forms a series of amphipathic a-helices (Ma et al, 1991;Riechmann and Meyerowitz, 1997b).…”

Section: Introductionmentioning

confidence: 99%

The K domain mediates heterodimerization of theArabidopsisfloral organ identity proteins, APETALA3 and PISTILLATA

2003

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SummaryMADS genes in plants encode key developmental regulators of vegetative and reproductive development. The majority of well-characterized plant MADS proteins contain two conserved domains, the DNA-binding MADS domain and the K domain. The K domain is predicted to form three amphipathic a-helices referred to as K1, K2, and K3. In this report, we define amino acids and subdomains important for heterodimerization between the two Arabidopsis floral organ identity MADS proteins APETALA3 (AP3) and PISTILLATA (PI). Analysis of mutants defective in dimerization demonstrates that K1, K2 and the region between K1 and K2 are critical for the strength of AP3/PI dimerization. The majority of the critical amino acids are hydrophobic indicating that the K domain mediates AP3/PI interaction primarily through hydrophobic interactions. Specially, K1 of AP3 and PI resembles a leucine zipper motif. Most mutants defective in AP3/PI heterodimerization in yeast exhibit partial floral organ identity function in transgenic Arabidopsis. Our results also indicate that the motif containing Asn-98 and specific charged residues in K1 (Glu-97 in PI and Arg-102 in AP3) are important for both the strength and specificity of AP3/PI heterodimer formation.

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“…Since the NLS and the DNA-binding domain or SRF are spatially distinct, SRF should act well as an adapter between the plasmid DNA and the importin nuclear import machinery. 28,29 In experiments using SRF-expressing CV1 transfectants, we routinely observed decreased nuclear localization of micro-injected plasmids, including the SV40 promoter/enhancer containing pCAT-control. While it is possible that the aberrant expression of SRF caused a down-regulation of general plasmid nuclear import, perhaps by transcriptionally regulating the expression of one or more transcription factors needed for SV40 DNA nuclear import, it is more likely that the decreased nuclear import of the pCAT-control plasmid was due to experimental variation.…”

Section: Discussionmentioning

confidence: 99%

Cell-specific nuclear import of plasmid DNA

et al. 1999

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One factor limiting the success of non-viral gene therapy smooth muscle cells, we have created a series of reporter vectors is the relative inability to target genes specifically plasmids that are expressed selectively in smooth muscle to a desired cell type. To address this limitation, we have cells. Moreover, when injected into the cytoplasm, plasbegun to develop cell-specific vectors whose specificity is mids containing portions of the SMGA promoter localize to at the level of the nuclear import of the plasmid DNA. We the nucleus of smooth muscle cells, but remain cytoplashave recently shown that nuclear import of plasmid DNA mic in fibroblasts and CV1 cells. In contrast, a similar plasis a sequence-specific event, requiring the SV40 enhancer, mid carrying the SV40 enhancer is transported into the a region known to bind to a number of general transcription nuclei of all cell types tested. Nuclear import of the SMGA factors (Dean DA, Exp Cell Res 1997; 230: 293). From promoter-containing plasmids could be achieved when the these studies we developed a model whereby transcription smooth muscle specific transcription factor SRF was factor(s) bind to the DNA in the cytoplasm to create a proexpressed in stably transfected CV1 cells, supporting our tein-DNA complex that can enter the nucleus using the model for the nuclear import of plasmids. Finally, these protein import machinery. Our model predicts that by using nuclear targeting sequences were also able to promote DNA elements containing binding sites for transcription facincreased gene expression in liposome-and polycationtors expressed in unique cell types, we should be able to transfected non-dividing cells in a cell-specific manner, create plasmids that target to the nucleus in a cell-specific similar to their nuclear import activity. These results promanner. Using the promoter from the smooth muscle vide proof of principle for the development of cell-specific gamma actin (SMGA) gene whose expression is limited to non-viral vectors for any desired cell type.

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Structure of serum response factor core bound to DNA

Cited by 320 publications

References 47 publications

Organization and Myogenic Restricted Expression of the Murine Serum Response Factor Gene

Organization and Myogenic Restricted Expression of the Murine Serum Response Factor Gene

The K domain mediates heterodimerization of theArabidopsisfloral organ identity proteins, APETALA3 and PISTILLATA

Cell-specific nuclear import of plasmid DNA

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