XXII. BIOCHEMICKÝ ZJAZD - Jesseniova lekárska fakulta

Posters
34.
PrEParaTION aND fUNCTIONal anaLYSIS of PHOSPHOrYLaTION
MUTaNT forMS of THE traNSCrIPTION faCTOr NFI
Gabriel Kollárovič, Miroslava Kretová, Peter Baráth and Katarína Luciaková
Laboratory of Molecular Biology, Cancer Research Institute, SAS, Bratislava
Nuclear factors I (NFI) are transcription factors and has been implicated in many cellular
processes such as embryonic development, cell differentiation and transformation
processes. In vertebrates, NFI proteins are encoded by four isogenes and exist
in multiple splice variant and can act both as activators and repressors. Recent data
from the functional proteomic studies identify NFIs as a substrate for ATM/ATR kinase
in response to DNA damage. Such genomic instability results in activation of a protein
cascade(s) that leads to the cell cycle arrest. Regarding our previous data on the role of
NFI in growth-regulated gene expression, a logical question arises whether NFI proteins
indeed play a role in the ATM/ATR signaling pathway. For this purpose specific mutations
in the ATM/ATR phosphorylation sites in the NFI was prepared to study its effects on
the expression of target genes. The serine to alanine point mutations were prepared by
PCR. Simultaneously restriction site was introduced for restriction enzyme SacI for rapid
screening of clones bearing the mutated forms of NFI.
Acknowledgements: This work was supported by VEGA grant No. 2/0074/08.
152 XXII. Biochemistry Congress, Martin