Transcription of the Lung-specific Surfactant Protein C Gene Is Mediated by Thyroid Transcription Factor 1

Kelly, Susan E.; Bachurski, Cindy J.; Burhans, Michael S.; Glasser, Stephan W.

doi:10.1074/jbc.271.12.6881

Cited by 229 publications

(176 citation statements)

References 29 publications

Supporting

Mentioning

163

Contrasting

Order By: Relevance

“…For transfection experiments, cells were seeded at 2x10 5 cells/well in six-well plates (TPP, Trasadingen, Switzerland) 24 h before transfection and transiently co-transfected using lipofectin (Invitrogen, Carlsbad, CA) with 3 μg of reporter plasmid, either SP-B luciferase [Yan et al, 1995] or SP-C luciferase plasmid [Kelly et al, 1996]; 2 μg of mock vector pcDNA3; and wild-type NKX2-1 or mutated NKX2-1 as specified in the figure legends. The cells were co-transfected with pRL-TK (Promega, Madison, WI) to normalize cell transfection efficiency.…”

Section: Cell Culture and Transactivation Experimentsmentioning

confidence: 99%

“…Nuclear proteins were further extracted using the NE-PER nuclear kit (Pierce, Rockford, IL). Two micrograms of nuclear protein extract were mixed with 40 fmol of biotinylated labeled SFTPB (SP-B-f1, [Bohinski et al, 1994]), SFTPC [Kelly et al, 1996], and TG (oligonucleotide C, [Santisteban et al, 1992]) probes (Sigma-Aldrich) derived from the corresponding promoters, as recommended by the manufacturer. For competition incubations, a 100-fold excess of cold probe was used.…”

Section: Emsamentioning

confidence: 99%

See 1 more Smart Citation

NKX2-1mutations leading to surfactant protein promoter dysregulation cause interstitial lung disease in “Brain-Lung-Thyroid Syndrome”

et al. 2010

View full text Add to dashboard Cite

NKX2-1 (NK2 homeobox 1) is a critical regulator of transcription for the surfactant protein (SP)-B and -C genes (SFTPB and SFTPC, respectively). We identified and functionally characterized two new de novo NKX2-1 mutations c.493C>T (p.R165W) and c.786_787del2 (p.L263fs) in infants with closely similar severe interstitial lung disease (ILD), hypotonia, and congenital hypothyroidism. Functional analyses using A549 and HeLa cells revealed that NKX2-1-p.L263fs induced neither SFTPB nor SFTPC promoter activation and had a dominant negative effect on wild-type (WT) NKX2-1. In contrast, NKX2-1-p.R165W activated SFTPC, to a significantly greater extent than did WT NKX2-1, while SFTPB activation was only significantly reduced in HeLa cells. In accordance with our in vitro data, we found decreased amounts of SP-B and SP-C by western blot in bronchoalveolar lavage fluid (patient with p.L263fs) and features of altered surfactant protein metabolism on lung histology (patient with NKX2-1-p.R165W). In conclusion, ILD in patients with NKX2-1 mutations was associated with altered surfactant protein metabolism, and both gain and loss of function of the mutated NKX2-1 genes on surfactant protein promoters were associated with ILD in "Brain-Lung-Thyroid syndrome".

show abstract

Section: Cell Culture and Transactivation Experimentsmentioning

confidence: 99%

Section: Emsamentioning

confidence: 99%

NKX2-1mutations leading to surfactant protein promoter dysregulation cause interstitial lung disease in “Brain-Lung-Thyroid Syndrome”

et al. 2010

View full text Add to dashboard Cite

show abstract

“…Restriction of SP-C expression to the lung epithelium is conferred by regulatory elements located within Ϫ215 bp of the human SP-C promoter (13). This region, which is highly homologous between humans and mice, contains conserved binding sites for thyroid transcription factor 1, a phosphorylated, homeodomain-containing member of the Nkx nuclear transcription factor family (TTF-1; also called Nkx2.1 and T-EBP; mouse genomic designation Titf1) (14). TTF-1 is expressed in the epithelium of the developing lung, thyroid, and in the central nervous system.…”

mentioning

confidence: 99%

Erm/Thyroid Transcription Factor 1 Interactions Modulate Surfactant Protein C Transcription

Lin

Perl

Shannon

2006

Journal of Biological Chemistry

View full text Add to dashboard Cite

Expression of surfactant protein C (SP-C), which is restricted to alveolar type II epithelial cells of the adult lung, is critically dependent on thyroid transcription factor 1 (TTF-1). In the present study we have demonstrated that Erm, a member of the Ets family of transcription factors, is expressed in the distal lung epithelium during development and is also restricted to alveolar type II cells in the adult. Erm was up-regulated by fibroblast growth factors (FGFs) in culture, and blocking FGF signaling inhibited Erm expression both in vivo and in vitro. The SP-C minimal promoter was found to contain two potential Ets binding sites, and electrophoretic mobility shift assays showed that two 20-bp wild-type oligonucleotides containing the 5-GGA(A/T)-3 Ets consensus binding motif were shifted by nuclear extracts from MLE15 cells. Co-transfection assays showed that Erm by itself had little effect on SP-C promoter activity but that Erm significantly enhanced TTF-1-mediated SP-C transcription. Mutation of one of the Ets binding sites reduced SP-C transcription to background levels, whereas mutation of the other site resulted in increased SP-C transcription. Protein-protein interactions between Erm and TTF-1 were demonstrated by mammalian two-hybrid assays and by co-immunoprecipitation assays. Mapping studies showed that the Ets domain of Erm and the combined N terminus and homeodomain of TTF-1 were critical for this interaction. Treatment of primary cultures of adult alveolar type II cells with siRNA targeting Erm diminished expression of both Erm and SP-C but had no effect on ␤-actin or GAPDH (glyceraldehyde-3-phosphate dehydrogenase). Taken together, these results demonstrate that Erm is involved in SP-C regulation, which results from an interaction with TTF-1.Mouse lung development begins on embryonic day 9.5 (E9.5) 2 as two evaginations from the floor of the foregut endoderm into mesenchyme derived from splanchnic mesoderm. After a series of dichotomous and lateral branchings that form the pulmonary tree, the most distal epithelium continues to expand and mature during late gestation and early postnatal life to give rise to mature alveoli. Two cell types constitute the alveolar epithelium, alveolar type I and type II cells. Type I cells have a highly attenuated morphology that is ideally suited for gas exchange (1), whereas the more cuboidal type II cells function in a number of capacities. These include serving as progenitor cells for type I cells, directionally transporting sodium from apical to basolateral cell surfaces to minimize alveolar fluid, producing molecules involved in innate host defense, and synthesizing and secreting pulmonary surfactant (2).Proximate tissue interactions between the epithelium and mesenchyme play a critical role in lung morphogenesis and differentiation. Tissue recombination studies have shown that distal lung epithelial differentiation is induced as a specific response to signals produced by lung mesenchyme (3, 4). Furthermore, tissue recombination experiments in which epitheli...

show abstract

“…The CF cuboidal cells of the alveoli were shown to possess characteristics of type II pneumocytes. For example, they exhibited immunoreactivity to TTF-1, a transcription factor that is known to specifically activate genes encoding constituents of surfactant (Bruno et al, 1995;Kelly et al, 1996). They also expressed CK 7, a protein that is found in the alveolar cells (Broers et al 1989;Iyonaga et al, 1997).…”

Section: Discussionmentioning

confidence: 99%

“…To characterize cuboidal alveolar cells, we investigated the expression of proteins evidenced in type II pneumocytes in healthy pulmonary tissue, namely cystic fibrosis transmembrane conductance regulator (McCray et al, 1993;Engelhardt et al, 1994), cytokeratin 7 (Broers et al 1989;Iyonaga et al, 1997), thyroid transcription factor-1 (Bruno et al, 1995;Kelly et al, 1996), as well as cytokeratin 5 a marker of basal and parabasal cells in normal human airway epithelium (Boers et al, 1998) and the Ki-67 antigen, a marker of proliferating cells (Gerdes et al, 1984). The immunohistochemical reactions showed that the alveolar cuboidal cells expressed cystic fibrosis transmembrane conductance regulator (Fig.…”

Section: Cytological Characterization Of Cf Alveoli Before Explantationmentioning

confidence: 99%

Growth of putative progenitors of type II pneumocytes in culture of human cystic fibrosis alveoli

Hollande

Cantet

Ratovo

et al. 2004

Biology of the Cell

View full text Add to dashboard Cite

Alveolar type II pneumocytes are thought to be progenitor cells capable of self-renewal and differentiation into type I pneumocytes. Nevertheless, the existence of an alveolar stem cell has been postulated. In lungs from patients with cystic fibrosis, the alveolar epithelium is damaged with ulceration and subsequent regeneration. We characterized alveolar modifications histologically and immunohistochemically in the pulmonary tissue of a patient homozygous for the DF508 mutation. Alveoli were of variable size and surrounded by an inflammatory infiltrate. They were lined by a continuous layer of cuboidal cells with very weak proliferative activity. These cells resembled type II pneumocytes. They expressed thyroid transcription factor-1, cystic fibrosis transmembrane conductance regulator, cytokeratin 7 and contained lamellar bodies. Weak expression of cytokeratin 5 considered to be a marker of progenitor cells of the bronchial and bronchiolar epithelium was detected. Explantation of this alveolar epithelium produced primary cultures and subcultures of epithelial cells that had acquired proliferative properties showing signs of dedifferentiation with a loss of lamellar bodies and a lack of expression of thyroid transcription factor-1. Persistence of the expression of cytokeratin 7 and a strong expression of cytokeratin 5 were observed. The culture conditions were thought to have circumvented the inhibition of proliferation observed in vivo due to the inflammatory peri-alveolar environment. They thus favored the multiplication of a population of cells co-expressing cytokeratin 5 and certain characteristics of type II pneumocytes. The presence of these cells of intermediate phenotype is indicative of the existence of immature precursors for type II pneumocytes.

show abstract

Transcription of the Lung-specific Surfactant Protein C Gene Is Mediated by Thyroid Transcription Factor 1

Cited by 229 publications

References 29 publications

NKX2-1mutations leading to surfactant protein promoter dysregulation cause interstitial lung disease in “Brain-Lung-Thyroid Syndrome”

NKX2-1mutations leading to surfactant protein promoter dysregulation cause interstitial lung disease in “Brain-Lung-Thyroid Syndrome”

Erm/Thyroid Transcription Factor 1 Interactions Modulate Surfactant Protein C Transcription

Growth of putative progenitors of type II pneumocytes in culture of human cystic fibrosis alveoli

Contact Info

Product

Resources

About