Recombinant Human ATF1

• Cat.No.: ATF1-27042TH
• Product Overview: Recombinant fusion protein, full length human ATF1 expressed by baculovirus in Sf9 insect cells using a N-terminal proprietary tag.

Specification

• Species: Human
• Tag: Non
• Description: This gene encodes an activating transcription factor, which belongs to the ATF subfamily and bZIP (basic-region leucine zipper) family. It influences cellular physiologic processes by regulating the expression of downstream target genes, which are related to growth, survival, and other cellular activities. This protein is phosphorylated at serine 63 in its kinase-inducible domain by serine/threonine kinases, cAMP-dependent protein kinase A, calmodulin-dependent protein kinase I/II, mitogen- and stress-activated protein kinase and cyclin-dependent kinase 3 (cdk-3). Its phosphorylation enhances its transactivation and transcriptional activities, and enhances cell transformation. Fusion of this gene and FUS on chromosome 16 or EWSR1 on chromosome 22 induced by translocation generates chimeric proteins in angiomatoid fibrous histiocytoma and clear cell sarcoma. This gene has a pseudogene on chromosome 6.
• Form: Liquid
• Storage buffer: Preservative: NoneConstituents: 25% Glycerol, 50mM Tris HCl, 150mM Sodium chloride, 0.25mM DTT, 0.1mM PMSF, pH 7.5
• Storage: Shipped on dry ice. Upon delivery aliquot and store at -80oC. Avoid freeze / thaw cycles.
• Sequence Similarities: Belongs to the bZIP family. ATF subfamily.Contains 1 bZIP domain.Contains 1 KID (kinase-inducible) domain.
• Full Length: Full L.

Gene Information

• Gene Name: ATF1 activating transcription factor 1 [ Homo sapiens ]
• Official Symbol: ATF1
• Synonyms: ATF1; activating transcription factor 1; cyclic AMP-dependent transcription factor ATF-1; TREB36
• Gene ID: 466
• mRNA Refseq: NM_005171
• Protein Refseq: NP_005162
• MIM: 123803
• Uniprot ID: P18846
• Chromosome Location: 12q13
• Pathway: Activated TLR4 signalling, organism-specific biosystem; CREB phosphorylation, organism-specific biosystem; EGFR1 Signaling Pathway, organism-specific biosystem; ErbB1 downstream signaling, organism-specific biosystem; HTLV-I infection, organism-specific biosystem
• Function: protein complex binding; protein heterodimerization activity; sequence-specific DNA binding; sequence-specific DNA binding transcription factor activity; sequence-specific distal enhancer binding RNA polymerase II transcription factor activity

Citation

CCR5AS lncRNA variation differentially regulates CCR5, influencing HIV disease outcome

Journal: Nature immunology    PubMed ID: 31209403    Data: 2019/4/27

Authors: Smita Kulkarni, Alexandra Lied, Mary Carrington

Article Snippet:Nuclear extracts prepared from HeLa and Jurkat cell lines via the CellLyticNuCLEAR extraction kit (Sigma-Aldrich) were stored at ?80°C until use.Nuclear extracts prepared from HeLa and Jurkat cell lines via the CellLyticNuCLEAR extraction kit (Sigma-Aldrich) were stored at ?80°C until use.. Lysate from HEK 293 transfected with recombinant human Atf1 (Creative BioMart) was used as a positive control.. Double-stranded DNA oligonucleotide probes incorporating the predicted ATF-1 binding sequence (Fwd: 5’- GTGAAAGCTTG TTAC G TA GGTAACCTTTTGT-3’; Rev: 5’- GGACAAAAGGTTACC TA C GTAA CAAGCTTT-3’) or the mutant sequence (Fwd: 5’- GTGAAAGCTTG TTAC A TA GGTAACCTTTTGT-3’; Rev: 5’- GGACAAAAGGTTACC TA T GTAA CAAGCTTT-3’) were synthesized and labelled using Klenow fragment of DNA polymerase I (Invitrogen) with [α? 32 P]deoxycytidine triphosphate (3000 Ci/mmol; PerkinElmer Life and Analytical Sciences).Double-stranded DNA oligonucleotide probes incorporating the predicted ATF-1 binding sequence (Fwd: 5’- GTGAAAGCTTG TTAC G TA GGTAACCTTTTGT-3’; Rev: 5’- GGACAAAAGGTTACC TA C GTAA CAAGCTTT-3’) or the mutant sequence (Fwd: 5’- GTGAAAGCTTG TTAC A TA GGTAACCTTTTGT-3’; Rev: 5’- GGACAAAAGGTTACC TA T GTAA CAAGCTTT-3’) were synthesized and labelled using Klenow fragment of DNA polymerase I (Invitrogen) with [α? 32 P]deoxycytidine triphosphate..

a , An electrophoretic mobility shift assay (EMSA) was performed with 32 P-labelled oligonucleotides containing the core binding sequence for ATF1 containing the A or G variant of rs2027820. Nuclear extracts of recombinant human ATF1-transfected HEK 293, or untransfected HeLa or Jurkat cells were incubated with each of the oligonucleotides (indicated as A or G) in the presence of a control antibody (Ctl for ATF1-transfected HEK 293 extracts was specific for Sp1; Ctl for HeLa and Jurkat extracts was nonspecific IgG) or ATF1-specific antibody (ATF1). The presence of a shift (marked Atf1) indicates that the oligonucleotide binds a protein contained in the corresponding nuclear extract. A supershift is observed when an anti-ATF1 antibody is added, but not when Sp1 antibody or control IgG is added. A single experiment was performed. b , A fragment of 1000 base pairs immediately upstream of the transcription start site of CCR5AS, and a 500 base pair intronic fragment containing the rs2027820A/G polymorphic site were cloned upstream and downstream of the firefly luciferase gene, respectively, in a PGL3-basic vector. The firefly luciferase constructs with basic vector (no promoter), control vector (SV40 promoter), IntA (rs2027820A), and IntG (rs rs2027820G) were transfected along with Renilla luciferase encoding vector in a 293T derivative cell line (Lenti-X). c , Luciferase activity was measured 24 hours post-transfection and relative activity of Firefly luciferase normalized to Renilla was determined as relative light units (RLU). The IntG vector showed significantly higher luciferase activity as compared to the IntA vector. The data represent three replicates in each group from two independent experiments experiments. The mean ±SE are depicted as horizontal and vertical bars for each group, respectively. Statistical significance was determined using non-parametric t test and two tailed p value is indicated.