HDGF and nucleolin complex formation depends on bcl-2 full length mRNA. (A) To investigate a possible involvement of bcl-2 mRNA in HDGF-nucleolin complex formation cells were treated with Paclitaxel and protein lysates were examined for nucleolin copurification as described above. Incubation with the bcl-2 mRNA destabilizing agent Paclitaxel led to a significant decrease of nucleolin copurification. (B + C) Different deletion constructs of bcl-2 mRNA were generated and cotransfected together with HDGFStrep-tag to investigate their influence on the copurification of nucleolin. The full-length bcl-2 mRNA (FL Bcl-2) had a positive whereas the coding sequence of bcl-2 (cds) had a negative effect on the amount of coprecipitated nucleolin. Interestingly, addition of 400 bp of the 3′UTR (ARE1) containing a known nucleolin binding motive was already sufficient to rescue the negative effect on the amount of precipitated nucleolin. Furthermore, beside the 3′UTR also the 5′UTR had a rescuing effect on the interaction.
Bremer et al. BMC Biochemistry 2013 14:2 doi:10.1186/1471-2091-14-2