[Analysis and identification of transcriptional repression domain of ETO].
OBJECTIVE
To further verify the transcriptional repression domains in ETO and their relationship with histone deacetylase (HDAC).


METHODS
Either of the ETO two zinc fingers was mutated respectively by site-directing mutagenesis. The truncation fragments of ETO were amplified by polymerase chain reaction (PCR) and cloned into eukaryotic expression plasmid pFA-CMV. By the means of DNA transfection and analysis of the transcription derived from the promoter of reporter gene, the transcriptional regulation domains of ETO was determined.


RESULTS
The expression plasmids carrying truncated ETO and ETO with point mutation at either zinc finger were successfully constructed. Two repression domains were found within ETO, which were located at two zinc finger motifs and 275 - 487 amino acid residues, respectively.


CONCLUSIONS
The transcription repression by ETO was mediated by two separated domains and closely associated with HDAC, which may be used as therapeutic target for acute myeloid leukemia M(2b).
Wang
Min
M
Department of Clinical Hematology, State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Diseases Hospital, CAMS and PUMC, Tianjin 300020, China.
Wang
Ling
L
Hao
Chang-lai
CL
Xing
Hai-yan
HY
Tang
Ke-jing
KJ
Wang
Jian-xiang
JX
D050676:Core Binding Factor Alpha 2 Subunit / Q000737:chemistry / Q000235:genetics; D015973:Gene Expression Regulation, Leukemic / Q000235:genetics*; D005822:Genetic Vectors; D006655:Histone Deacetylases / Q000502:physiology; D006801:Humans; D066298:In Vitro Techniques; D015470:Leukemia, Myeloid, Acute / Q000201:enzymology / Q000235:genetics*; D015514:Oncogene Proteins, Fusion / Q000737:chemistry / Q000235:genetics; D000075142:RUNX1 Translocation Partner 1 Protein; D014158:Transcription, Genetic; D014162:Transfection; D016335:Zinc Fingers / Q000235:genetics
C107844:AML1-ETO fusion protein, human; D050676:Core Binding Factor Alpha 2 Subunit; D015514:Oncogene Proteins, Fusion; D000075142:RUNX1 Translocation Partner 1 Protein; D006655:Histone Deacetylases