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• W1978548976 abstract "IRF-1 is a tumor suppressor protein that activates gene expression from a range of promoters in response to stimuli spanning viral infection to DNA damage. Studies on the post-translational regulation of IRF-1 have been hampered by a lack of suitable biochemical tools capable of targeting the endogenous protein. In this study, phage display technology was used to develop a monoclonal nanobody targeting the C-terminal Mf1 domain (residues 301–325) of IRF-1. Intracellular expression of the nanobody demonstrated that the transcriptional activity of IRF-1 is constrained by the Mf1 domain as nanobody binding gave an increase in expression from IRF-1-responsive promoters of up to 8-fold. Furthermore, Mf1-directed nanobodies have revealed an unexpected function for this domain in limiting the rate at which the IRF-1 protein is degraded. Thus, the increase in IRF-1 transcriptional activity observed on nanobody binding is accompanied by a significant reduction in the half-life of the protein. In support of the data obtained using nanobodies, a single point mutation (P325A) involving the C-terminal residue of IRF-1 has been identified, which results in greater transcriptional activity and a significant increase in the rate of degradation. The results presented here support a role for the Mf1 domain in limiting both IRF-1-dependent transcription and the rate of IRF-1 turnover. In addition, the data highlight a route for activation of downstream genes in the IRF-1 tumor suppressor pathway using biologics. IRF-1 is a tumor suppressor protein that activates gene expression from a range of promoters in response to stimuli spanning viral infection to DNA damage. Studies on the post-translational regulation of IRF-1 have been hampered by a lack of suitable biochemical tools capable of targeting the endogenous protein. In this study, phage display technology was used to develop a monoclonal nanobody targeting the C-terminal Mf1 domain (residues 301–325) of IRF-1. Intracellular expression of the nanobody demonstrated that the transcriptional activity of IRF-1 is constrained by the Mf1 domain as nanobody binding gave an increase in expression from IRF-1-responsive promoters of up to 8-fold. Furthermore, Mf1-directed nanobodies have revealed an unexpected function for this domain in limiting the rate at which the IRF-1 protein is degraded. Thus, the increase in IRF-1 transcriptional activity observed on nanobody binding is accompanied by a significant reduction in the half-life of the protein. In support of the data obtained using nanobodies, a single point mutation (P325A) involving the C-terminal residue of IRF-1 has been identified, which results in greater transcriptional activity and a significant increase in the rate of degradation. The results presented here support a role for the Mf1 domain in limiting both IRF-1-dependent transcription and the rate of IRF-1 turnover. In addition, the data highlight a route for activation of downstream genes in the IRF-1 tumor suppressor pathway using biologics. IntroductionThe skipping of exon 3 and/or 2 seen in transcripts for the IRF-1 tumor suppressor has been linked to the development of human hemopoietic malignancies, such as leukemia and myelodysplastic syndrome (1Boultwood J. Fidler C. Lewis S. MacCarthy A. Sheridan H. Kelly S. Oscier D. Buckle V.J. Wainscoat J.S. Blood. 1993; 82: 2611-2616Crossref PubMed Google Scholar, 2Green W.B. Slovak M.L. Chen I.M. Pallavicini M. Hecht J.L. Willman C.L. Leukemia. 1999; 13: 1960-1971Crossref PubMed Scopus (54) Google Scholar, 3Willman C.L. Sever C.E. Pallavicini M.G. Harada H. Tanaka N. Slovak M.L. Yamamoto H. Harada K. Meeker T.C. List A.F. et al.Science. 1993; 259: 968-971Crossref PubMed Scopus (379) Google Scholar). More recently, the loss of exons 7–9 in various combinations has been reported in cervical cancer, and this leads to the generation of C-terminally truncated IRF-1 that can compete with the wild-type protein for DNA binding. The C-terminal mutants have longer half-lives and, unlike wild-type IRF-1, can be expressed in a cell cycle independent manner (4Lee E.J. Jo M. Park J. Zhang W. Lee J.H. Biochem. Biophys. Res. Commun. 2006; 347: 882-888Crossref PubMed Scopus (31) Google Scholar). Understanding the function of the Mf1 domain of IRF-1, a regulatory subdomain that is located at the C terminus of the protein, could therefore increase our current knowledge of the role that truncated proteins play in tumor progression.The extreme C-terminal region of IRF-1 (Mf1 domain; amino acids 301–325) is a regulatory domain that plays a role in both positive and negative modulation of target gene expression (5Eckert M. Meek S.E. Ball K.L. J. Biol. Chem. 2006; 281: 23092-23102Abstract Full Text Full Text PDF PubMed Scopus (14) Google Scholar). In addition, the Mf1 domain plays a role in determining the rate of IRF-1 protein degradation, with removal of the C terminus leading to a significant increase in the half-life of the protein (6Pion E. Narayan V. Eckert M. Ball K.L. Cell. Signal. 2009; 21: 1479-1487Crossref PubMed Scopus (22) Google Scholar, 7Nakagawa K. Yokosawa H. Eur. J. Biochem. 2000; 267: 1680-1686Crossref PubMed Scopus (73) Google Scholar). Furthermore, binding of members of the Hsp70 family of molecular chaperones to the Mf1 domain appears to be critical for the normal function and regulation of IRF-1 (8Narayan V. Eckert M. Zylicz A. Zylicz M. Ball K.L. J. Biol. Chem. 2009; 284: 25889-25899Abstract Full Text Full Text PDF PubMed Scopus (19) Google Scholar). Thus previous studies have pointed to a potentially important role for the Mf1 domain of IRF-1 in its homeostatic regulation.The studies described above, by necessity, relied on the use of exogenous IRF-1 mutant proteins; therefore, we do not know whether the Mf1 domain is rate-limiting for IRF-1-mediated gene activation within the context of the endogenous protein. To address this issue, we have used antibody phage display to screen for scFv 3The abbreviations used are: scFvsingle chain variable fragmentNi-NTAnickel-nitrilotriacetic acidTRAILtumor necrosis factor-related apoptosis-inducing ligandISREinterferon-stimulated response elementEGFPenhanced GFPPKRprotein kinase RNA-activated. antibody fragments (9Smith G.P. Science. 1985; 228: 1315-1317Crossref PubMed Scopus (3040) Google Scholar, 10McCafferty J. Griffiths A.D. Winter G. Chiswell D.J. Nature. 1990; 348: 552-554Crossref PubMed Scopus (1904) Google Scholar) to the Mf1 domain. The introduction of scFv nanobodies into a cell system showed that the Mf1 domain was rate-limiting for IRF-1-mediated transcription under normal cellular conditions, providing good evidence that IRF-1 transcriptional function is subject to post-translational regulation. The data also highlight an unexpected role for the extreme C terminus in the negative regulation of IRF-1 turnover.DISCUSSIONData are presented suggesting that the Mf1 domain of IRF-1 limits its ability to activate transcription and that this negative regulation can be relieved using nanobodies that bind to the Mf1 domain or by mutating the C-terminal residue of the protein (Pro325). In addition, the data highlight a possible role for the Mf1 domain in coordinating a link between the rate of IRF-1 degradation and its transcriptional activity. Previous studies have suggested that IRF-1 activity is regulated primarily at the level of transcription as IRF-1 steady state levels increase in response to agents such as interferon and DNA damage, which induce its transcriptional activity (3Willman C.L. Sever C.E. Pallavicini M.G. Harada H. Tanaka N. Slovak M.L. Yamamoto H. Harada K. Meeker T.C. List A.F. et al.Science. 1993; 259: 968-971Crossref PubMed Scopus (379) Google Scholar, 25Romeo G. Fiorucci G. Chiantore M.V. Percario Z.A. Vannucchi S. Affabris E. J. Interferon Cytokine Res. 2002; 22: 39-47Crossref PubMed Scopus (101) Google Scholar). However, we demonstrate here that IRF-1 is also subject to post-translational regulation; it is held in a latent or partially active state, and activity can be induced in the absence of an increase in its steady state levels.The Mf1 domain is a 25-amino acid region in the extreme C terminus of IRF-1, which appears to be a “hot spot” for the regulation of both its function and turnover. Thus, the Mf1 domain has been shown to house an LXXLL coregulator signature motif (amino acids 306–310), which is essential for IRF-1-mediated repression of Cdk2 (5Eckert M. Meek S.E. Ball K.L. J. Biol. Chem. 2006; 281: 23092-23102Abstract Full Text Full Text PDF PubMed Scopus (14) Google Scholar) and a binding site for members of the Hsp70 family of molecular chaperones that play a role in maintaining the normal cellular function of IRF-1 (8Narayan V. Eckert M. Zylicz A. Zylicz M. Ball K.L. J. Biol. Chem. 2009; 284: 25889-25899Abstract Full Text Full Text PDF PubMed Scopus (19) Google Scholar). The Mf1 domain lies within a region originally defined as an enhancer region (amino acids 257–325) required for maximal IRF-1 transcriptional activity (26Kirchhoff S. Oumard A. Nourbakhsh M. Levi B.Z. Hauser H. Eur. J. Biochem. 2000; 267: 6753-6761Crossref PubMed Scopus (27) Google Scholar) that most likely functions through the recruitment of coactivators such as p300 (27Dornan D. Eckert M. Wallace M. Shimizu H. Ramsay E. Hupp T.R. Ball K.L. Mol. Cell. Biol. 2004; 24: 10083-10098Crossref PubMed Scopus (58) Google Scholar). More detailed analysis of the IRF-1 C terminus has revealed that removal of the last 25 residues, rather than the entire enhancer domain, does not compromise its ability to activate gene expression. Rather a ΔC-terminal 25-residue IRF-1 truncation mutant displayed an increase in intrinsic transactivation potential when compared with the full-length protein (Fig. 5) (5Eckert M. Meek S.E. Ball K.L. J. Biol. Chem. 2006; 281: 23092-23102Abstract Full Text Full Text PDF PubMed Scopus (14) Google Scholar). This suggests that the extreme C terminus, later named multifunctional-1 (Mf1) domain (8Narayan V. Eckert M. Zylicz A. Zylicz M. Ball K.L. J. Biol. Chem. 2009; 284: 25889-25899Abstract Full Text Full Text PDF PubMed Scopus (19) Google Scholar), might be a negative regulator of IRF-1 activity. As these studies relied on the use of transiently expressed IRF-1 mutant proteins, it has not been possible to determine the effect of the Mf1 domain on IRF-1 as a transcriptional activator under normal cellular conditions. By developing a nanobody that can bind to both endogenous and exogenous IRF-1, we have been able to demonstrate that the Mf1 domain is normally rate-limiting for IRF-1-mediated gene expression. The data presented in this study are in good agreement with our previous observations using Ala scanning and a series of truncation mutants to narrow down the possible negative regulatory domain (5Eckert M. Meek S.E. Ball K.L. J. Biol. Chem. 2006; 281: 23092-23102Abstract Full Text Full Text PDF PubMed Scopus (14) Google Scholar) to amino acids 311–317. As the scFv3 interaction with IRF-1 requires both Tyr311 and Ser317 (Fig. 4), its activation of IRF-1-mediated gene expression lends strong support to the idea that these residues include a negative regulatory motif (5Eckert M. Meek S.E. Ball K.L. J. Biol. Chem. 2006; 281: 23092-23102Abstract Full Text Full Text PDF PubMed Scopus (14) Google Scholar).Previous studies have shown that single chain nanobodies can be used to modulate the function of their target antigens in a cellular environment by, for example, altering the intracellular localization of the target proteins (18Cossins A.J. Harrison S. Popplewell A.G. Gore M.G. Protein Expr. Purif. 2007; 51: 253-259Crossref PubMed Scopus (18) Google Scholar, 28Beerli R.R. Wels W. Hynes N.E. J. Biol. Chem. 1994; 269: 23931-23936Abstract Full Text PDF PubMed Google Scholar, 29Zhou P. Goldstein S. Devadas K. Tewari D. Notkins A.L. J. Immunol. 1998; 160: 1489-1496PubMed Google Scholar, 30Tellez C. Jean D. Bar-Eli M. Methods. 2004; 34: 233-239Crossref PubMed Scopus (2) Google Scholar), neutralizing enzyme activity (31Paz K. Brennan L.A. Iacolina M. Doody J. Hadari Y.R. Zhu Z. Mol. Cancer Ther. 2005; 4: 1801-1809Crossref PubMed Scopus (42) Google Scholar), disrupting normal protein-protein interactions (32Visintin M. Melchionna T. Cannistraci I. Cattaneo A. J. Biotechnol. 2008; 135: 1-15Crossref PubMed Scopus (26) Google Scholar, 33Riley C.J. Engelhardt K.P. Saldanha J.W. Qi W. Cooke L.S. Zhu Y. Narayan S.T. Shakalya K. Croce K.D. Georgiev I.G. Nagle R.B. Garewal H. Von Hoff D.D. Mahadevan D. Cancer Res. 2009; 69: 1933-1940Crossref PubMed Scopus (33) Google Scholar, 34Cohen P.A. Mani J.C. Lane D.P. Oncogene. 1998; 17: 2445-2456Crossref PubMed Scopus (52) Google Scholar, 35Bai J. Sui J. Zhu R.Y. Tallarico A.S. Gennari F. Zhang D. Marasco W.A. J. Biol. Chem. 2003; 278: 1433-1442Abstract Full Text Full Text PDF PubMed Scopus (71) Google Scholar), or affecting DNA-protein interactions (32Visintin M. Melchionna T. Cannistraci I. Cattaneo A. J. Biotechnol. 2008; 135: 1-15Crossref PubMed Scopus (26) Google Scholar). In this study, the scFv3 nanobody was able to activate IRF-1-mediated transcription between 4- and 8-fold making it a powerful tool to study the functions of the Mf1 domain in a cell system. Conventional monoclonal antibodies and nanobodies have been used previously to study a negative regulatory domain in p53. Monoclonals and scFv, which bind to the 421 epitope, have been used to explore the role of the p53 C-terminal domain in negative regulation of its DNA binding and transcriptional activity (36Hupp T.R. Lane D.P. Curr. Biol. 1994; 4: 865-875Abstract Full Text Full Text PDF PubMed Scopus (301) Google Scholar, 37Hupp T.R. Meek D.W. Midgley C.A. Lane D.P. Cell. 1992; 71: 875-886Abstract Full Text PDF PubMed Scopus (859) Google Scholar, 38Hupp T.R. Meek D.W. Midgley C.A. Lane D.P. Nucleic Acids Res. 1993; 21: 3167-3174Crossref PubMed Scopus (185) Google Scholar). In addition, 421 antibody-based scFvs have been suggested as a way to partially reactivate mutant forms of the p53 protein in cells (39Caron de Fromentel C. Gruel N. Venot C. Debussche L. Conseiller E. Dureuil C. Teillaud J.L. Tocque B. Bracco L. Oncogene. 1999; 18: 551-557Crossref PubMed Scopus (69) Google Scholar). The mechanism of p53 activation by 421 antibodies is through an increase in the sequence-specific DNA binding activity of the protein. In this study, we found no evidence for an increased DNA binding activity of IRF-1 in the presence of scFv3 (Fig. 3e) nor was there any obvious difference in the subcellular localization of IRF-1 when bound to the activating nanobody (Fig. 7a). Nanobody binding may impact on the conformation/dynamics of the IRF-1 structure or attenuate inhibition mediated via an intramolecular interaction. The IRF-1-related protein IRF-3 is subject to this type of regulation. In the case of IRF-3, autoinhibition occurs due to an interaction between the C terminus and a region from the N-terminal domain (40Lin R. Mamane Y. Hiscott J. Mol. Cell. Biol. 1999; 19: 2465-2474Crossref PubMed Scopus (268) Google Scholar), and this process is regulated by C-terminal phosphorylation (41Yang H. Lin C.H. Ma G. Orr M. Baffi M.O. Wathelet M.G. Eur. J. Biochem. 2002; 269: 6142-6151Crossref PubMed Scopus (38) Google Scholar). However, structural information is only available for the DNA binding domain of IRF-1 (42Escalante C.R. Yie J. Thanos D. Aggarwal A.K. Nature. 1998; 391: 103-106Crossref PubMed Scopus (313) Google Scholar), and how the Mf1 domain relates to the overall tertiary structure of the protein remains to be determined. It is also possible that the Mf1 domain can act as an allosteric modulatory site in response to C-terminal binding proteins. To date, few IRF-1 interactions have been documented, and the number of proteins known to interact with the C terminus is even more limited. However, Hsp70 family members have recently been identified as physiologically relevant Mf1 interacting factors that impact on IRF-1 localization, transcriptional activity, and its rate of degradation (8Narayan V. Eckert M. Zylicz A. Zylicz M. Ball K.L. J. Biol. Chem. 2009; 284: 25889-25899Abstract Full Text Full Text PDF PubMed Scopus (19) Google Scholar) suggesting that the activity of IRF-1 is likely to be modulated by agents that mimic or disrupt Mf1 protein-protein interactions.Increased IRF-1 degradation upon scFv3 binding (Fig. 7, b–d) seems contrary to our previous observation that the Mf1 domain is required for efficient IRF-1 turnover (6Pion E. Narayan V. Eckert M. Ball K.L. Cell. Signal. 2009; 21: 1479-1487Crossref PubMed Scopus (22) Google Scholar). However, the Pro325 IRF-1 mutant protein displays a similar property as it has a reduced t0.5 compared with the wild-type protein (Fig. 8b). As nanobody epitope mapping (Fig. 4) shows that scFv3 also has a strong requirement for Pro325, its effect on degradation seems unlikely to be an artifact. The Mf1 domain therefore appears to contain elements that are both required for efficient degradation of IRF-1 (6Pion E. Narayan V. Eckert M. Ball K.L. Cell. Signal. 2009; 21: 1479-1487Crossref PubMed Scopus (22) Google Scholar) and that prevent the rate of degradation from being too rapid (FIGURE 7, FIGURE 8), i.e. the Mf1 can act like a rheostat “fine-tuning” IRF-1 turnover dependent on cellular conditions.Both scFv3-bound IRF-1 and P325A IRF-1 have increased transactivation activity, as well as an increased rate of degradation, making it interesting to speculate that these two Mf1 domain functions may be linked. It is increasingly clear that both the proteolytic and nonproteolytic functions of the proteasome are required for correct regulation of the transcriptional machinery, with evidence of both the 19 S and 26 S proteasomes being associated with chromatin, components of the basal transcription machinery, and/or various transcription factors (43Muratani M. Tansey W.P. Nat. Rev. Mol. Cell Biol. 2003; 4: 192-201Crossref PubMed Scopus (668) Google Scholar, 44Lipford J.R. Deshaies R.J. Nat. Cell Biol. 2003; 5: 845-850Crossref PubMed Scopus (156) Google Scholar, 45Reid J. Svejstrup J.Q. J. Biol. Chem. 2004; 279: 29875-29878Abstract Full Text Full Text PDF PubMed Scopus (45) Google Scholar, 46Somesh B.P. Reid J. Liu W.F. S⊘gaard T.M. Erdjument-Bromage H. Tempst P. Svejstrup J.Q. Cell. 2005; 121: 913-923Abstract Full Text Full Text PDF PubMed Scopus (178) Google Scholar, 47Dhananjayan S.C. Ismail A. Nawaz Z. Essays Biochem. 2005; 41: 69-80Crossref PubMed Google Scholar, 48Ransom M. Williams S.K. Dechassa M.L. Das C. Linger J. Adkins M. Liu C. Bartholomew B. Tyler J.K. J. Biol. Chem. 2009; 284: 23461-23471Abstract Full Text Full Text PDF PubMed Scopus (52) Google Scholar). The precise mechanism(s) linking transcriptional activation with protein degradation remain unclear. However, evidence suggests roles for proteasome-mediated degradation in establishing limits for transcription, promoting the exchange of transcription factors on chromatin, and stimulating multiple rounds of transcription initiation (49Gillette T.G. Gonzalez F. Delahodde A. Johnston S.A. Kodadek T. Proc. Natl. Acad. Sci. U.S.A. 2004; 101: 5904-5909Crossref PubMed Scopus (130) Google Scholar, 50Collins G.A. Tansey W.P. Curr. Opin. Genet. Dev. 2006; 16: 197-202Crossref PubMed Scopus (213) Google Scholar, 51Kang Z. Pirskanen A. Jänne O.A. Palvimo J.J. J. Biol. Chem. 2002; 277: 48366-48371Abstract Full Text Full Text PDF PubMed Scopus (164) Google Scholar, 52Saccani S. Marazzi I. Beg A.A. Natoli G. J. Exp. Med. 2004; 200: 107-113Crossref PubMed Scopus (213) Google Scholar, 53Tanaka T. Grusby M.J. Kaisho T. Nat. Immunol. 2007; 8: 584-591Crossref PubMed Scopus (230) Google Scholar, 54Ostendorff H.P. Peirano R.I. Peters M.A. Schlüter A. Bossenz M. Scheffner M. Bach I. Nature. 2002; 416: 99-103Crossref PubMed Scopus (137) Google Scholar). It will therefore be of interest to determine exactly how changes in the half-life of IRF-1 relate to its function as a transcription factor. IntroductionThe skipping of exon 3 and/or 2 seen in transcripts for the IRF-1 tumor suppressor has been linked to the development of human hemopoietic malignancies, such as leukemia and myelodysplastic syndrome (1Boultwood J. Fidler C. Lewis S. MacCarthy A. Sheridan H. Kelly S. Oscier D. Buckle V.J. Wainscoat J.S. Blood. 1993; 82: 2611-2616Crossref PubMed Google Scholar, 2Green W.B. Slovak M.L. Chen I.M. Pallavicini M. Hecht J.L. Willman C.L. Leukemia. 1999; 13: 1960-1971Crossref PubMed Scopus (54) Google Scholar, 3Willman C.L. Sever C.E. Pallavicini M.G. Harada H. Tanaka N. Slovak M.L. Yamamoto H. Harada K. Meeker T.C. List A.F. et al.Science. 1993; 259: 968-971Crossref PubMed Scopus (379) Google Scholar). More recently, the loss of exons 7–9 in various combinations has been reported in cervical cancer, and this leads to the generation of C-terminally truncated IRF-1 that can compete with the wild-type protein for DNA binding. The C-terminal mutants have longer half-lives and, unlike wild-type IRF-1, can be expressed in a cell cycle independent manner (4Lee E.J. Jo M. Park J. Zhang W. Lee J.H. Biochem. Biophys. Res. Commun. 2006; 347: 882-888Crossref PubMed Scopus (31) Google Scholar). Understanding the function of the Mf1 domain of IRF-1, a regulatory subdomain that is located at the C terminus of the protein, could therefore increase our current knowledge of the role that truncated proteins play in tumor progression.The extreme C-terminal region of IRF-1 (Mf1 domain; amino acids 301–325) is a regulatory domain that plays a role in both positive and negative modulation of target gene expression (5Eckert M. Meek S.E. Ball K.L. J. Biol. Chem. 2006; 281: 23092-23102Abstract Full Text Full Text PDF PubMed Scopus (14) Google Scholar). In addition, the Mf1 domain plays a role in determining the rate of IRF-1 protein degradation, with removal of the C terminus leading to a significant increase in the half-life of the protein (6Pion E. Narayan V. Eckert M. Ball K.L. Cell. Signal. 2009; 21: 1479-1487Crossref PubMed Scopus (22) Google Scholar, 7Nakagawa K. Yokosawa H. Eur. J. Biochem. 2000; 267: 1680-1686Crossref PubMed Scopus (73) Google Scholar). Furthermore, binding of members of the Hsp70 family of molecular chaperones to the Mf1 domain appears to be critical for the normal function and regulation of IRF-1 (8Narayan V. Eckert M. Zylicz A. Zylicz M. Ball K.L. J. Biol. Chem. 2009; 284: 25889-25899Abstract Full Text Full Text PDF PubMed Scopus (19) Google Scholar). Thus previous studies have pointed to a potentially important role for the Mf1 domain of IRF-1 in its homeostatic regulation.The studies described above, by necessity, relied on the use of exogenous IRF-1 mutant proteins; therefore, we do not know whether the Mf1 domain is rate-limiting for IRF-1-mediated gene activation within the context of the endogenous protein. To address this issue, we have used antibody phage display to screen for scFv 3The abbreviations used are: scFvsingle chain variable fragmentNi-NTAnickel-nitrilotriacetic acidTRAILtumor necrosis factor-related apoptosis-inducing ligandISREinterferon-stimulated response elementEGFPenhanced GFPPKRprotein kinase RNA-activated. antibody fragments (9Smith G.P. Science. 1985; 228: 1315-1317Crossref PubMed Scopus (3040) Google Scholar, 10McCafferty J. Griffiths A.D. Winter G. Chiswell D.J. Nature. 1990; 348: 552-554Crossref PubMed Scopus (1904) Google Scholar) to the Mf1 domain. The introduction of scFv nanobodies into a cell system showed that the Mf1 domain was rate-limiting for IRF-1-mediated transcription under normal cellular conditions, providing good evidence that IRF-1 transcriptional function is subject to post-translational regulation. The data also highlight an unexpected role for the extreme C terminus in the negative regulation of IRF-1 turnover." @default.
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• W1978548976 title "Intracellular Activation of Interferon Regulatory Factor-1 by Nanobodies to the Multifunctional (Mf1) Domain" @default.
• W1978548976 cites W130153342 @default.
• W1978548976 cites W1490624051 @default.
• W1978548976 cites W1504721248 @default.
• W1978548976 cites W1550304352 @default.
• W1978548976 cites W1583341340 @default.
• W1978548976 cites W1975793508 @default.
• W1978548976 cites W1995953017 @default.
• W1978548976 cites W1996790111 @default.
• W1978548976 cites W2000771665 @default.
• W1978548976 cites W2000919345 @default.
• W1978548976 cites W2001684550 @default.
• W1978548976 cites W2003236156 @default.
• W1978548976 cites W2005546492 @default.
• W1978548976 cites W2005635949 @default.
• W1978548976 cites W2006551649 @default.
• W1978548976 cites W2011261569 @default.
• W1978548976 cites W2021067723 @default.
• W1978548976 cites W2021998328 @default.
• W1978548976 cites W2024853038 @default.
• W1978548976 cites W2032131702 @default.
• W1978548976 cites W2036032993 @default.
• W1978548976 cites W2038834815 @default.
• W1978548976 cites W2041086320 @default.
• W1978548976 cites W2042498352 @default.
• W1978548976 cites W2048233287 @default.
• W1978548976 cites W2053472341 @default.
• W1978548976 cites W2055952888 @default.
• W1978548976 cites W2060584995 @default.
• W1978548976 cites W2064555088 @default.
• W1978548976 cites W2070360032 @default.
• W1978548976 cites W2071290275 @default.
• W1978548976 cites W2081115041 @default.
• W1978548976 cites W2086374403 @default.
• W1978548976 cites W2090345852 @default.
• W1978548976 cites W2091406431 @default.
• W1978548976 cites W2095696891 @default.
• W1978548976 cites W2097372287 @default.
• W1978548976 cites W2120929471 @default.
• W1978548976 cites W2127144150 @default.
• W1978548976 cites W2127494207 @default.
• W1978548976 cites W2147881852 @default.
• W1978548976 cites W2154161526 @default.
• W1978548976 cites W2155176680 @default.
• W1978548976 cites W2160213398 @default.
• W1978548976 cites W2160894282 @default.
• W1978548976 cites W2161197634 @default.
• W1978548976 cites W2167065653 @default.
• W1978548976 cites W2169000984 @default.
• W1978548976 cites W2315753416 @default.
• W1978548976 cites W2317640291 @default.
• W1978548976 cites W2356498392 @default.
• W1978548976 cites W2398665740 @default.
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