SUMOylation regulates the nuclear mobility of CREB binding protein and its association with nuclear bodies in live cells

Autor: David M. Heery, Karin B. Kindle, Colm M. Ryan, Hilary M. Collins
Rok vydání: 2010
Předmět:
CBP
CREB binding protein
SUMO-1 Protein
Biophysics
SUMO protein
HDAC
histone deacetylase
macromolecular substances
Biology
CBP
environment and public health
Biochemistry
Article
Cell Line
03 medical and health sciences
Nuclear bodies
0302 clinical medicine
PML
promyelocytic leukaemia
Live cell imaging
Transcription (biology)
Chlorocebus aethiops
Animals
Humans
FRAP
fluorescence recovery after photo-bleaching
CREB-binding protein
Molecular Biology
Sequence Deletion
030304 developmental biology
Cell Nucleus
0303 health sciences
PML
HEK 293 cells
TDG
thymine DNA glycosylase
Promoter
AML1
Cell Biology
Cell cycle
CREB-Binding Protein
YFP/GFP
yellow/green fluorescent protein
Molecular biology
SUMOylation
Chromatin
Cell biology
CRD
cell cycle repression domain
SUMO
small ubiquitin like modifier
COS Cells
FRAP
biology.protein
AML
acute myeloid leukaemia
030217 neurology & neurosurgery
Zdroj: Biochemical and Biophysical Research Communications
ISSN: 0006-291X
DOI: 10.1016/j.bbrc.2009.12.040
Popis: The lysine acetyltransferase CREB binding protein (CBP) is required for chromatin modification and transcription at many gene promoters. In fixed cells, a large proportion of CBP colocalises to PML or nuclear bodies. Using live cell imaging, we show here that YFP-tagged CBP expressed in HEK293 cells undergoes gradual accumulation in nuclear bodies, some of which are mobile and migrate towards the nuclear envelope. Deletion of a short lysine-rich domain that contains the major SUMO acceptor sites of CBP abrogated its ability to be SUMO modified, and prevented its association with endogenous SUMO-1/PML speckles in vivo. This SUMO-defective CBP showed enhanced ability to co-activate AML1-mediated transcription. Deletion mapping revealed that the SUMO-modified region was not sufficient for targeting CBP to PML bodies, as C-terminally truncated mutants containing this domain showed a strong reduction in accumulation at PML bodies. Fluorescence recovery after photo-bleaching (FRAP) experiments revealed that YFP–CBPΔ998–1087 had a retarded recovery time in the nucleus, as compared to YFP–CBP. These results indicate that SUMOylation regulates CBP function by influencing its shuttling between nuclear bodies and chromatin microenvironments.
Databáze: OpenAIRE
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