@article {10.3844/ajbbsp.2016.86.94,
article_type = {journal},
title = {Is RNA Binding in the Rossmann Fold Essential for GAPDH Intranuclear Functions?},
author = {Krynetskaia, Natalia and Phadke, Manali and Krynetskiy, Evgeny},
volume = {12},
number = {2},
year = {2016},
month = {Jun},
pages = {86-94},
doi = {10.3844/ajbbsp.2016.86.94},
url = {https://thescipub.com/abstract/ajbbsp.2016.86.94},
abstract = {In addition to its prominent enzymatic activity, GAPDH is an enigmatic component of multiple unrelated biochemical entities. In this study, we explored a series of mutated GAPDH and fusion EGFP-GAPDH polypeptides and compared nuclear accumulation, intranuclear mobility and RNA binding properties of the wild type and variant GAPDH proteins. Our results revealed that RNA binding to T99I-mutated GAPDH with non-functional NAD+ binding center occurred outside the Rossmann fold. At the cellular level, wild type and mutated EGFP-GAPDH demonstrated distinct intranuclear localization in unstressed cells versus cells exposed to genotoxic stress. Wild type EGFP-GAPDH protein localized in the cytoplasm of untreated cells and accumulated in the nucleus following araC treatment (11.8±2.72% vs. 27.4±4.28% nuclear EGFP-GAPDH, % of total EGFP-GAPDH, p = 0.0007). Mutated T99I EGFP-GAPDH accumulated at high level in the nuclei of untreated and araC-treated cells (34.3±8.49% vs. 41.3±16.0% nuclear EGFP-GAPDH, % of total EGFP-GAPDH, p = 0.21). Mutated T99I EGFP-GAPDH lost its ability to form tight interactions with intranuclear macromolecules. After araC treatment, immobile fraction (1-Mf) of wild type EGFP-GAPDH in the nuclei of SW48-297 cells was three times higher (0.75±0.127 vs. 0.26±0.133%, p},
journal = {American Journal of Biochemistry and Biotechnology},
publisher = {Science Publications}
}