SUMOylation of DRIL1 directs its transcriptional activity towards leukocyte lineage-specific genes.
DRIL1 is an ARID family transcription factor that can immortalize primary mouse fibroblasts, bypass RAS(V12)-induced cellular senescence and collaborate with RAS(V12) or MYC in mediating oncogenic transformation. It also activates immunoglobulin heavy chain transcription and engages in heterodimer formation with E2F to stimulate E2F-dependent transcription. Little, however, is known about the regulation of DRIL1 activity. Recently, DRIL1 was found to interact with the SUMO-conjugating enzyme Ubc9, but the functional relevance of this association has not been assessed. Here, we show that DRIL1 is sumoylated both in vitro and in vivo at lysine 398. Moreover, we provide evidence that PIASy functions as a specific SUMO E3-ligase for DRIL1 and promotes its sumoylation both in vitro and in vivo. Furthermore, consistent with the subnuclear localization of PIASy in the Matrix-Associated Region (MAR), SUMO-modified DRIL1 species are found exclusively in the MAR fraction. This post-translational modification interferes neither with the subcellular localization nor the DNA-binding activity of the protein. In contrast, DRIL1 sumoylation impairs its interaction with E2F1 in vitro and modifies its transcriptional activity in vivo, driving transcription of subset of genes regulating leukocyte fate. Taken together, these results identify sumoylation as a novel post-translational modification of DRIL1 that represents an important mechanism for targeting and modulating DRIL1 transcriptional activity.
Department of Molecular Genetics, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
Alexandre
A
Prieur
Karim
K
Nacerddine
Maarten
M
van Lohuizen
Daniel S
DS
Peeper
D000818:Animals; D002460:Cell Line; D019070:Cell Lineage / Q000235:genetics; D002478:Cells, Cultured; D004268:DNA-Binding Proteins; D006801:Humans; D007962:Leukocytes / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D009857:Oncogenes; D025842:SUMO-1 Protein / Q000378:metabolism*; D015534:Trans-Activators / Q000737:chemistry / Q000378:metabolism*; D014157:Transcription Factors; D014158:Transcription, Genetic / Q000235:genetics*; D014162:Transfection
C490798:ARID3A protein, human; D004268:DNA-Binding Proteins; D025842:SUMO-1 Protein; D015534:Trans-Activators; D014157:Transcription Factors