The C-terminal domain of the nuclear factor I-B2 isoform is glycosylated and transactivates the WAP gene in the JEG-3 cells.
The transcription factor nuclear factor I (NFI) has been shown previously both in vivo and in vitro to be involved in the cooperative regulation of whey acidic protein (WAP) gene transcription along with the glucocorticoid receptor and STAT5. In addition, one of the specific NFI isoforms, NFI-B2, was demonstrated in transient co-transfection experiments in JEG cells, which lack endogenous NFI, to be preferentially involved in the cooperative regulation of WAP gene expression. A comparison of the DNA-binding specificities of the different NFI isoforms only partially explained their differential ability to activate the WAP gene transcription. Here, we analyzed the transactivation regions of two NFI isoforms by making chimeric proteins between the NFI-A and B isoforms. Though, their DNA-binding specificities were not altered as compared to the corresponding wild-type transcription factors, the C-terminal region of the NFI-B isoform was shown to preferentially activate WAP gene transcription in cooperation with GR and STAT5 in transient co-transfection assays in JEG-3 cells. Furthermore, determination of serine and threonine-specific glycosylation (O-linked N-acetylglucosamine) of the C-terminus of the NFI-B isoform suggested that the secondary modification by O-GlcNAc might play a role in the cooperative regulation of WAP gene transcription by NFI-B2 and STAT5.
Mukhopadhyay
Sudit S
SS
Department of Molecular and Cancer Genetics, M.D. Anderson Cancer Center, Houston, TX 77030, USA. suditmukhopadhy@yahoo.com
Rosen
Jeffrey M
JM
D000117:Acetylglucosamine / Q000737:chemistry; D045744:Cell Line, Tumor; D006031:Glycosylation; D006801:Humans; D008894:Milk Proteins / Q000737:chemistry / Q000235:genetics*; D051839:NFI Transcription Factors / Q000737:chemistry*; D020033:Protein Isoforms; D017434:Protein Structure, Tertiary; D011965:Receptors, Glucocorticoid / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D050799:STAT5 Transcription Factor / Q000737:chemistry; D013912:Threonine / Q000737:chemistry; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D025521:Tumor Suppressor Proteins
D008894:Milk Proteins; D051839:NFI Transcription Factors; D020033:Protein Isoforms; D011965:Receptors, Glucocorticoid; D011993:Recombinant Fusion Proteins; D050799:STAT5 Transcription Factor; C494092:STAT5A protein, human; D025521:Tumor Suppressor Proteins; C024471:whey acidic proteins; D013912:Threonine; D000117:Acetylglucosamine