| First Author | Kang DH | Year | 2013 |
| Journal | Int J Mol Med | Volume | 32 |
| Issue | 5 | Pages | 1187-94 |
| PubMed ID | 24026233 | Mgi Jnum | J:294363 |
| Mgi Id | MGI:6456067 | Doi | 10.3892/ijmm.2013.1488 |
| Citation | Kang DH, et al. (2013) Novel function of the poly(c)-binding protein alpha-CP2 as a transcriptional activator that binds to single-stranded DNA sequences. Int J Mol Med 32(5):1187-94 |
| abstractText | alpha-complex protein 2 (alpha-CP2) is known as an RNA-binding protein that interacts in a sequence-specific manner with single-stranded polycytosine [poly(C)]. This protein is involved in various post-transcriptional regulations, such as mRNA stabilization and translational regulation. In this study, the full-length mouse alpha-CP2 gene was expressed in an insoluble form with an N-terminal histidine tag in Escherichia coli and purified for homogeneity using affinity column chromatography. Its identity was confirmed using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. Recombinant alpha-CP2 was expressed and refolded. The protein folding conditions for denatured alpha-CP2 were optimized. DNA and RNA electrophoretic mobility shift assays demonstrated that the recombinant alpha-CP2 is capable of binding to both single-stranded DNA and RNA poly(C) sequences. Furthermore, plasmids expressing alpha-CP2 activated the expression of a luciferase reporter when co-transfected with a single-stranded (pGL-SS) construct containing a poly(C) sequence. To our knowledge, this study demonstrates for the first time that alpha-CP2 functions as a transcriptional activator by binding to a single-stranded poly(C) sequence. |
