Revision as of 19:32, 26 September 2024

ATF2

Usage and Biology

ATF2 belongs to the ATF/CREB family and regulates genes involved in cell growth, stress responses and apoptosis. The ATF-2 protein is DNA-binding that binds to cyclic AMP-response elements (CREs), thereby forming a homodimer or heterodimer with c-Jun. It then stimulates CRE-dependent transcription (Kawasaki et al. , 2000)

We employed ATF2 as a transcription factor in our cell-based beta-lactam-sensor, which in contrast to CcpA (K5317014) or GraR (K5317015) originates from an eukaryotic background, to transfer the PknB-detected signal into reporter gene expression.

Cloning

Theoretical Part Design

The ATF2 gene was synthesized, and the gene sequence was explicity chose from its cDNA to exclude intons and shorten the gene sequence inserted into the final plasmid for characterization (K5317021).

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 21
Illegal EcoRI site found at 283
Illegal XbaI site found at 324
Illegal PstI site found at 731
Illegal PstI site found at 1180
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 21
Illegal EcoRI site found at 283
Illegal PstI site found at 731
Illegal PstI site found at 1180
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 21
Illegal EcoRI site found at 283
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 21
Illegal EcoRI site found at 283
Illegal XbaI site found at 324
Illegal PstI site found at 731
Illegal PstI site found at 1180
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 21
Illegal EcoRI site found at 283
Illegal XbaI site found at 324
Illegal PstI site found at 731
Illegal PstI site found at 1180
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI.rc site found at 1031

Characterization

The functionality of ATF2 in our cell-based sensor was assessed by analysing its general expression after transfection in HEK293T cells and assessing its localization by fusion with the reporter gene mRuby2 (K5317001). For results please visit the registry entry K5317021.

References

Kirsch, K., Zeke, A., Tőke, O., Sok, P., Sethi, A., Sebő, A., Kumar, G. S., Egri, P., Póti, Á. L., Gooley, P., Peti, W., Bento, I., Alexa, A., & Reményi, A. (2020). Co-regulation of the transcription controlling ATF2 phosphoswitch by JNK and p38. Nature Communications, 11(1), 5769. https://doi.org/10.1038/s41467-020-19582-3

@@ Line 5: / Line 5: @@
 ===Usage and Biology===
-ATF2 belongs to the ATF/CREB family and regulates genes involved in cell growth, stress responses and apoptosis. With the addition of the ATF2 gene, this plasmid enables the study of transcriptional regulation of ATF2 (Kirsch ''et al.'', 2020) and its phosphorylation by PknB, making it important for research into signaling pathways related to cell stress and survival.
+ATF2 belongs to the ATF/CREB family and regulates genes involved in cell growth, stress responses and apoptosis. The ATF-2 protein is DNA-binding that binds to cyclic AMP-response elements (CREs), thereby forming a homodimer or heterodimer with c-Jun. It then stimulates CRE-dependent transcription (Kawasaki  ''et al. '', 2000)
 We employed ATF2 as a transcription factor in our cell-based beta-lactam-sensor, which in contrast to CcpA (<span class="plainlinks">[https://parts.igem.org/Part:BBa_K5317014 K5317014]</span>) or GraR (<span class="plainlinks">[https://parts.igem.org/Part:BBa_K5317015 K5317015]</span>) originates from an eukaryotic background, to transfer the PknB-detected signal into reporter gene expression.
 =Cloning=