The FHA domain determines Drosophila chk2/Mnk localization to key mitotic structures and is essential for early embryonic DNA damage responses

Saeko Takada, Eric R. Collins, Kayo Kurahashi

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

Abstract

DNA damage responses, including mitotic centrosome inactivation, cell-cycle delay in mitosis, and nuclear dropping from embryo cortex, maintain genome integrity in syncytial Drosophila embryos. A conserved signaling kinase, Chk2, known as Mnk/Loki, is essential for the responses. Here we demonstrate that functional EGFP-Mnk expressed from a transgene localizes to the nucleus, centrosomes, interkinetochore/centromere region, midbody, and pseudocleavage furrows without DNA damage and in addition forms numerous foci/aggregates on mitotic chromosomes upon DNA damage. We expressed EGFP-tagged Mnk deletion or point mutation variants and investigated domain functions of Mnk in vivo. A triple mutation in the phosphopeptide-binding site of the forkhead-associated (FHA) domain disrupted normal Mnk localization except to the nucleus. The mutation also disrupted Mnk foci formation on chromosomes upon DNA damage. FHA mutations and deletion of the SQ/TQ-cluster domain (SCD) abolished Mnk transphosphorylations and autophosphorylations, indicative of kinase activation after DNA damage. A potent NLS was found at the C-terminus, which is required for normal Mnk function. We propose that the FHA domain in Mnk plays essential dual functions in mediating embryonic DNA damage responses by means of its phosphopeptide-binding ability: activating Mnk in the nucleus upon DNA damage and recruiting Mnk to multiple subcellular structures independently of DNA damage.

Original languageEnglish (US)
Pages (from-to)1811-1828
Number of pages18
JournalMolecular biology of the cell
Volume26
Issue number10
DOIs
StatePublished - May 15 2015

Fingerprint Dive into the research topics of 'The FHA domain determines Drosophila chk2/Mnk localization to key mitotic structures and is essential for early embryonic DNA damage responses'. Together they form a unique fingerprint.

Cite this