Interaction between interferon consensus sequence-binding protein and COP9/signalosome subunit CSN2 (Trip15). A possible link between interferon regulatory factor signaling and the COP9/signalosome.
Interferon consensus sequence-binding protein (ICSBP) is a member of the interferon regulatory factors (IRF) that has a pivotal role in mediating resistance to pathogenic infections in mice and in promoting the differentiation of myeloid cells. ICSBP exerts some of its transcriptional activities via association with other factors that enable its binding to a variety of promoters containing DNA composite elements. These interactions are mediated through a specific COOH-terminal domain termed IAD (IRF association domain). To gain a broader insight of the capacity of ICSBP to interact with other factors, yeast two-hybrid screens were performed using ICSBP-IAD as a bait against a B-cell cDNA library. Trip15 was identified as a specific interacting factor with ICSBP in yeast cells, which was also confirmed by in vitro glutathione S-transferase pull-down assays and by coimmunoprecipitation studies in COS7 cells. Trip15 was recently identified as a component of the COP9/signalosome (CSN) complex composed of eight evolutionary conserved subunits and thus termed CSN2. This complex has a role in cell-signaling processes, which is manifested by its associated novel kinase activity and by the involvement of its subunits in regulating multiple cell-signaling pathways and cell-cycle progression. We show that in vitro association of ICSBP with the CSN leads to phosphorylation of ICSBP at a unique serine residue within its IAD. The phosphorylated residue is essential for efficient association with IRF-1 and thus for the repressor activity of ICSBP exerted on IRF-1. This suggests that the CSN has a role in integrating incoming signals that affect the transcriptional activity of ICSBP.
Cohen
H
H
Department of Food Engineering and Biotechnology, Technion, Haifa 32000, Israel.
Azriel
A
A
Cohen
T
T
Meraro
D
D
Hashmueli
S
S
Bech-Otschir
D
D
Kraft
R
R
Dubiel
W
W
Levi
B Z
BZ
D016475:3T3 Cells; D000409:Alanine / Q000737:chemistry; D000818:Animals; D001402:B-Lymphocytes / Q000378:metabolism; D015152:Blotting, Northern; D000075686:COP9 Signalosome Complex; D019556:COS Cells; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism*; D004247:DNA / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015723:Gene Library; D005982:Glutathione Transferase / Q000378:metabolism; D018922:HL-60 Cells; D006367:HeLa Cells; D006801:Humans; D050835:Interferon Regulatory Factors; D051379:Mice; D008954:Models, Biological; D046912:Multiprotein Complexes; D016297:Mutagenesis, Site-Directed; D009687:Nuclear Proteins; D010447:Peptide Hydrolases; D010766:Phosphorylation; D010957:Plasmids / Q000378:metabolism; D011233:Precipitin Tests; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D011988:Receptors, Thyroid Hormone; D012097:Repressor Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D012694:Serine / Q000737:chemistry; D015398:Signal Transduction; D014157:Transcription Factors; D014158:Transcription, Genetic; D020798:Two-Hybrid System Techniques; D000099185:Interferon Regulatory Factor-8
D000409:Alanine; D000075686:COP9 Signalosome Complex; D002352:Carrier Proteins; D004247:DNA; D018076:DNA, Complementary; D005982:Glutathione Transferase; D050835:Interferon Regulatory Factors; D046912:Multiprotein Complexes; D009687:Nuclear Proteins; D010447:Peptide Hydrolases; D011506:Proteins; D012333:RNA, Messenger; D011988:Receptors, Thyroid Hormone; D012097:Repressor Proteins; D012694:Serine; D014157:Transcription Factors; D000099185:Interferon Regulatory Factor-8; C118919:Cops2 protein, mouse