The interferon-inducible 15-kDa ubiquitin homolog conjugates to intracellular proteins

• Published in: The Journal of biological chemistry Vol. 267; no. 11; pp. 7806 - 7813
• Main Authors: LOEB, K. R, HAAS, A. L
• Format: Journal Article
• Language: English
• Published: Bethesda, MD American Society for Biochemistry and Molecular Biology 15.04.1992
• Subjects: A549 cells; Amino Acid Sequence; Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Blotting, Western; Cattle; Cytokines; Electrophoresis, Polyacrylamide Gel; Fundamental and applied biological sciences. Psychology; Humans; induction; interferon; Interferons - pharmacology; Miscellaneous; Molecular Sequence Data; Nuclear Proteins - genetics; Nuclear Proteins - metabolism; Protein Binding; Proteins; Proteins - metabolism; Rabbits; Sequence Homology, Nucleic Acid; Tumor Cells, Cultured; ubiquitin cross-reactive protein; Ubiquitins - analogs & derivatives; Ubiquitins - genetics; Ubiquitins - metabolism; Proteins; Ubiquitin; Induction; Molecular association; Immunoblotting assay; Homology; Interferon; Gene expression
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/s0021-9258(18)42585-9

We have previously identified a 15-kDa interferon-induced protein that is recognized by affinity-purified rabbit polyclonal antibodies against ubiquitin (Haas, A. L., Ahrens, P., Bright, P. M., and Ankel, H. (1987) J. Biol. Chem. 262, 11315-11323). This ubiquitin cross-reactive protein (UCRP) possesses significant homology to a tandem diubiquitin sequence. Since the biological effects of ubiquitin arise from its covalent ligation to intracellular target proteins, we hypothesized that the multiple cellular responses to inteferon are mediated in part by an analogous conjugation pathway for UCRP. Rabbit polyclonal antibodies specific for UCRP were prepared against homogeneous recombinant protein. Affinity-purified anti-UCRP antibodies detected the induction of UCRP in interferon-beta-treated A549 cells and recognized a group of high molecular weight UCRP conjugates on immunoblots of sodium dodecyl sulfate-polyacrylamide gel electrophoresis-resolved cell extracts. Both free and conjugated UCRP are constitutively present at low levels in untreated cells, suggesting a role for UCRP ligation in normal cellular regulation, and significantly accumulate following interferon treatment. The temporal induction of free UCRP following interferon treatment preceded a delayed increase in UCRP conjugates. Treatment of A549 cells with type I interferons (alpha and beta) strongly induced the expression of free and conjugated UCRP, whereas the response to type II interferon (gamma) was significantly less. A survey of selected cultured cell lines showed differential induction of free versus conjugated UCRP pools in response to interferon. Interferon-beta treatment of A549, MG63, and U937 cells induced high levels of both free and conjugated UCRP, whereas only free UCRP levels increased in Daudi, Namalwa, and K562 cells. These results confirm that UCRP represents a functional ubiquitin homolog participating in a parallel pathway of post-translational ligation and provides a novel mechanism for the response of susceptible cells to the effects of interferon exposure.