Cloning and characterization of the mouse polyamine-modulatedfactor-1 (mPMF-1) gene: an alternatively spliced homologue of the human transcription factor

2001 ◽  
Vol 359 (2) ◽  
pp. 387-392 ◽  
Author(s):  
Yanlin WANG ◽  
Wendy DEVEREUX ◽  
Patrick M. WOSTER ◽  
Robert A. CASERO
Keyword(s):  
Amino Acids ◽  
Transcription Factor ◽  
Transcriptional Activation ◽  
Splice Variants ◽  
Coiled Coil ◽  
Human Protein ◽  
Related Factor ◽  
Polyamine Analogues ◽  
Coiled Coil Region ◽  
Alternatively Spliced

The natural polyamines and their analogues have been implicated in transcriptional regulation of specific genes. Human polyamine-modulated factor-1 (hPMF-1) was the first polyamine-responsive transcription factor identified. Here the mouse homologue of the hPMF-1 gene is described. Interestingly, the mouse gene (mPMF-1) codes for two alternatively spliced mRNAs. Both of the mouse splice variants, mPMF-1S and mPMF-1L, possess C-terminal coiled-coil domains nearly identical to that found in hPMF-1 and are highly homologous with the human protein. The C-terminal coiled-coil structure is necessary for transcriptional activation. However, the shorter protein, mPMF-1S, does not contain an N-terminal coiled-coil region as do both hPMF-1 and the longer mPMF-1L. mPMF-1L mRNA codes for a protein of 202 amino acids, 37 amino acids longer than the human protein. By contrast, mPMF-1S codes for only 133 amino acids, as a result of two exons being omitted compared with mPMF-1L. Both mouse transcription factors can interact with Nrf-2 (nuclear factor-E2-related factor 2), the normal partner of hPMF-1, substantiating the importance of the C-terminal coiled-coil region responsible for this interaction. Finally, the expression of mPMF-1 is induced when mouse M1 myeloid leukaemia cells are exposed to polyamine analogues, suggesting control similar to that observed for the hPMF-1.

Characterization of the interaction between the transcription factors human polyamine modulated factor (PMF-1) and NF-E2-related factor 2 (Nrf-2) in the transcriptional regulation of the spermidine/spermine N1-acetyltransferase (SSAT) gene

2001 ◽  
Vol 355 (1) ◽  
pp. 45-49 ◽  
Author(s):  
Yanlin WANG ◽  
Wendy DEVEREUX ◽  
Tracy Murray STEWART ◽  
Robert A. CASERO
Keyword(s):  
Transcriptional Regulation ◽  
Transcription Factors ◽  
Leucine Zipper ◽  
Coiled Coil ◽  
Protein Family ◽  
Related Factor ◽  
Polyamine Analogues ◽  
Coiled Coil Region

Polyamines and polyamine analogues have been demonstrated to modulate the transcription of various genes. Spermidine/spermine N1-acetyltransferase (SSAT) is transcriptionally regulated through the interaction of at least two trans-acting transcription factors, NF-E2-related factor 2 (Nrf-2) and PMF-1 (polyamine modulated factor-1). Nrf-2has previously been shown to regulate transcription of other genes through interactions between its C-terminal leucine zipper and the leucine-zipper region of other members of the small Maf protein family (the term ‘Maf’ is derived from musculoaponeurotic-fibrosarcoma virus). Here it is demonstrated that the interaction between Nrf-2 and PMF-1 is mediated through the binding of the leucine-zipper region of Nrf-2 and a C-terminal coiled-coil region of PMF-1 that does not contain a leucine zipper. Mutations that interrupt either the leucine zipper of Nrf-2 or the coiled-coil region of PMF-1 are demonstrated to alter the ability of these factors to interact, thus their ability to regulate the transcription of the SSAT gene is lost.

Identification of Proteins That Interact with the Central Coiled-Coil Region of the Human Protein Kinase NEK1†

Biochemistry ◽  
2003 ◽  
Vol 42 (51) ◽  
pp. 15369-15376 ◽  
Author(s):  
Marcelo J. Surpili ◽  
Tatiana M. Delben ◽  
Jörg Kobarg
Keyword(s):  
Protein Kinase ◽  
Coiled Coil ◽  
Human Protein ◽  
Human Protein Kinase ◽  
Coiled Coil Region

scholarly journals Positive and Negative Regulation of the Transforming Growth Factor β/Activin Target Gene goosecoid by the TFII-I Family of Transcription Factors

2005 ◽  
Vol 25 (16) ◽  
pp. 7144-7157 ◽  
Author(s):  
Manching Ku ◽  
Sergei Y. Sokol ◽  
Jack Wu ◽  
Maria Isabel Tussie-Luna ◽  
Ananda L. Roy ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Growth Factor ◽  
Transcriptional Activation ◽  
Target Gene ◽  
Transforming Growth Factor ◽  
Transforming Growth Factor Β ◽  
Related Factor ◽  
P19 Cells ◽  
Binding Factor ◽  
Opposing Effects

ABSTRACT Goosecoid (Gsc) is a homeodomain-containing transcription factor present in a wide variety of vertebrate species and known to regulate formation and patterning of embryos. Here we show that in embryonic carcinoma P19 cells, the transcription factor TFII-I forms a complex with Smad2 upon transforming growth factor β (TGFβ)/activin stimulation, is recruited to the distal element (DE) of the Gsc promoter, and activates Gsc transcription. Downregulation of endogenous TFII-I by small inhibitory RNA in P19 cells abolishes the TGFβ-mediated induction of Gsc. Similarly, Xenopus embryos with endogenous TFII-I expression downregulated by injection of TFII-I-specific antisense oligonucleotides exhibit decreased Gsc expression. Unlike TFII-I, the related factor BEN (binding factor for early enhancer) is constitutively recruited to the distal element in the absence of TGFβ/activin signaling and is replaced by the TFII-I/Smad2 complex upon TGFβ/activin stimulation. Overexpression of BEN in P19 cells represses the TGFβ-mediated transcriptional activation of Gsc. These results suggest a model in which TFII-I family proteins have opposing effects in the regulation of the Gsc gene in response to a TGFβ/activin signal.

scholarly journals Endogenous p53 protein generated from wild-type alternatively spliced p53 RNA in mouse epidermal cells

1994 ◽  
Vol 14 (3) ◽  
pp. 1698-1708
Author(s):  
M F Kulesz-Martin ◽  
B Lisafeld ◽  
H Huang ◽  
N D Kisiel ◽  
L Lee
Keyword(s):  
Cell Cycle ◽  
Amino Acids ◽  
Transcriptional Activation ◽  
Cultured Cells ◽  
P53 Protein ◽  
Epidermal Cells ◽  
Wild Type ◽  
Mouse Tissues ◽  
Alternatively Spliced ◽  
In Cells

We previously demonstrated that a wild-type alternatively spliced p53 (p53as) RNA exists in mouse cultured cells and normal mouse tissues at approximately 25 to 33% of the level of the major p53 RNA form. The alternative RNA transcript is 96 nucleotides longer than the major transcript as a result of alternative splicing of intron 10 sequences. The protein expected to be generated from the p53as transcript is 9 amino acids shorter than the major p53 protein and has 17 different amino acids at the carboxyl terminus. We report here that p53as protein exists in nontransformed and malignant epidermal cells and is localized to the nucleus. In addition, p53as protein is preferentially expressed during the G2 phase of the cell cycle and in cells with greater than G2 DNA content compared with the major p53 protein, which is preferentially expressed in G1. The p53as immunoreactivity is elevated and shifted to the G1 phase of the cell cycle following actinomycin D treatment of nontransformed cells but not malignant cells. In view of the dimerization and tetramerization of p53 protein which may be necessary for its DNA binding and transcriptional activation activities, the presence of p53as protein in cells has important implications for understanding the physiological function(s) of the p53 gene.

β1PIX, the PAK-interacting exchange factor, requires localization via a coiled-coil region to promote microvillus-like structures and membrane ruffles

2001 ◽  
Vol 114 (23) ◽  
pp. 4239-4251 ◽  
Author(s):  
Cheng-Gee Koh ◽  
Ed Manser ◽  
Zhou-Shen Zhao ◽  
Chee-Peng Ng ◽  
Louis Lim
Keyword(s):  
Splice Variants ◽  
Coiled Coil ◽  
Cell Periphery ◽  
Nucleotide Exchange ◽  
Dimerization Domain ◽  
Intracellular Location ◽  
Exchange Factor ◽  
C Terminus ◽  
Nucleotide Exchange Factor ◽  
Coiled Coil Region

PIX is a Rho-family guanine nucleotide exchange factor that binds PAK. We previously described two isoforms of PIX that differ in their N termini. Here, we report the identification of a new splice variant of βPIX, designated β2PIX, that is the dominant species in brain and that lacks the region of ∼120 residues with predicted coiled-coil structure at the C terminus of β1PIX. Instead, β2PIX contains a serine-rich C terminus. To determine whether these splice variants differ in their cellular function, we studied the effect of expressing these proteins in HeLa cells. We found that the coiled-coil region plays a key role in the localization of β1PIX to the cell periphery and is also responsible for PIX dimerization. Overexpression of β1, but not β2PIX, drives formation of membrane ruffles and microvillus-like structures (via activation of Rac1 and Cdc42, respectively), indicating that its function requires localized activation of these GTPases. Thus, β1PIX, like other RhoGEFs, exerts specific morphological functions that are dependent on its intracellular location and are mediated by its C-terminal dimerization domain.

scholarly journals Amino Acids Control Mammalian Gene Transcription: Activating Transcription Factor 2 Is Essential for the Amino Acid Responsiveness of the CHOP Promoter

2000 ◽  
Vol 20 (19) ◽  
pp. 7192-7204 ◽  
Author(s):  
Alain Bruhat ◽  
Céline Jousse ◽  
Valérie Carraro ◽  
Andreas M. Reimold ◽  
Marc Ferrara ◽  
...  
Keyword(s):  
Gene Expression ◽  
Amino Acids ◽  
Transcription Factor ◽  
Amino Acid ◽  
Transcriptional Activation ◽  
Dominant Negative Mutant ◽  
Transient Expression Assay ◽  
Control Of Gene Expression ◽  
Mammalian Gene ◽  
Activating Transcription Factor

ABSTRACT In mammals, plasma concentration of amino acids is affected by nutritional or pathological conditions. It has been well established that nutrients, and particularly amino acids, are involved in the control of gene expression. Here we examined the molecular mechanisms involved in the regulation ofCHOP (a CCAAT/enhancer-binding protein [C/EBP]-related gene) expression upon amino acid limitation. We have previously shown that regulation of CHOP mRNA expression by amino acid concentration has both transcriptional and posttranscriptional components. We report the analysis ofcis- and trans-acting elements involved in the transcriptional activation of the human CHOPgene by leucine starvation. Using a transient expression assay, we show that a cis-positive element is essential for amino acid regulation of the CHOP promoter. This sequence is the first described that can regulate a basal promoter in response to starvation for several individual amino acids and therefore can be called an amino acid response element (AARE). In addition, we show that the CHOP AARE is related to C/EBP and ATF/CRE binding sites and binds in vitro the activating transcription factor 2 (ATF-2) in starved and unstarved conditions. Using ATF-2-deficient mouse embryonic fibroblasts and an ATF-2-dominant negative mutant, we demonstrate that expression of this transcription factor is essential for the transcriptional activation of CHOP by leucine starvation. Altogether, these results suggest that ATF-2 may be a member of a cascade of molecular events by which the cellular concentration of amino acids can regulate mammalian gene expression.

Differentiation-Dependent Interactions Between Runx-1 and Fli-1 during Megakaryocyte Development

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 279-279
Author(s):  
Hui Huang ◽  
Ming Yu ◽  
Tyler B Moran ◽  
Nathan Tu ◽  
Thomas E Akie ◽  
...  
Keyword(s):  
Amino Acids ◽  
Transcription Factor ◽  
Dna Binding ◽  
Transcriptional Activation ◽  
Gel Filtration ◽  
Cell Types ◽  
Myelogenous Leukemia ◽  
Activation Domains ◽  
Platelet Disorder ◽  
Familial Platelet Disorder

Abstract The transcription factor Runx-1 is required for the ontogeny of all definitive hematopoiesis, and plays a specific role in megakaryopoiesis during later stages of development. Germline mutations in Runx-1 cause Familial Platelet Disorder with Propensity to Develop AML (FPD/AML), and acquired mutations occur in a subset of patients with myelodysplastic syndrome (MDS) and acute myelogenous leukemia. Although many of the reported Runx-1 mutations affect DNA binding and/or its interaction with the cofactor CBF-beta, other mutations occur outside of these binding regions and have unknown mechanistic effects. In this study, we purified Runx-1 containing multiprotein complexes from murine megakaryocytic cells in order to identify potential novel Runx-1 associated factors whose interaction may be altered by Runx-1 mutations. Here we report the identification of the key megakaryocyte ets transcription factor Fli- 1 as a direct Runx-1 binding partner. This interaction involves the negative regulatory DNA binding and activation domains of Runx-1 (amino acids 179–370), and a region around the Ets DNA binding region of Fli-1 (amino acids 281–361). The interaction is lost in the MDS- associated Y254X Runx-1 mutation. We also show that Runx-1 and Fli-1 co-occupy the c-mpl promoter in primary megakaryocytes and act synergistically in transcriptional reporter assays. Interestingly, the interaction between Runx-1 and Fli- 1 occurs in murine L8057 megakaryoblastic cells only after they have been induced to differentiate, even though both proteins are expressed abundantly in uninduced cells. The interaction correlates with assembly of a large multiprotein complex that also includes the key megakaryocyte transcription factor GATA-1 and its cofactor Friend of GATA-1 (FOG- 1) based on gel filtration chromatography experiments. Furthermore, we show that Fli-1 from this large complex lacks phosphorylation of a specific residue that is phosphorylated on non-complexed Fli-1. Mutation of this site to aspartic acid, which mimics constitutive phosphorylation, disrupts the interaction between Fli-1 and Runx-1 and abrogates their transcriptional synergy. We propose that dephosphorylation of Fli-1 is a key event in the transcriptional activation of megakaryocyte terminal maturation by facilitating the assembly of a RUNX-1/FLI-1/GATA-1/FOG-1 enhancesome complex. These findings have implications for the differentiation of other cell types where interactions between Runx and ets family proteins occur.

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