Selective RNA Binding by a Single CCCH Zinc-Binding Domain from Nup475 (Tristetraprolin)

Michel, Sarah L. J.; Guerrerio, Anthony L.; Berg, Jeremy M.

doi:10.1021/bi034073h

Cited by 59 publications

(83 citation statements)

References 19 publications

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“…Alternatively, ZAP may recognize as yet unidentified short motifs or secondary structures, with the affinity for any such single motif being too low to support sustained binding; the clustering of these sequences would synergistically increase the affinity. The latter speculation is supported by the facts that there are four CCCH-type zinc finger motifs, each one of which is presumably an RNA binding unit (16,17), and that disruption of any of the zinc fingers reduced ZAP's activity to some extent (Fig. 4).…”

Section: Discussionmentioning

confidence: 90%

The Zinc Finger Antiviral Protein Directly Binds to Specific Viral mRNAs through the CCCH Zinc Finger Motifs

Guo

Carroll

MacDonald

et al. 2004

J Virol

232

342

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The zinc finger antiviral protein (ZAP) is a recently isolated host antiviral factor. It specifically inhibits the replication of Moloney murine leukemia virus (MLV) and Sindbis virus (SIN) by preventing the accumulation of viral RNA in the cytoplasm. For this report, we mapped the viral sequences that are sensitive to ZAP inhibition. The viral sequences were cloned into a luciferase reporter and analyzed for the ability to mediate ZAP-dependent destabilization of the reporter. The sensitive sequence in MLV was mapped to the 3 long terminal repeat; the sensitive sequences in SIN were mapped to multiple fragments. The fragment of SIN that displayed the highest destabilizing activity was further analyzed by deletion mutagenesis for the minimal sequence that retained the activity. This led to the identification of a fragment of 653 nucleotides. Any further deletion of this fragment resulted in significantly lower activity. We provide evidence that ZAP directly binds to the active but not the inactive fragments. The CCCH zinc finger motifs of ZAP play important roles in RNA binding and antiviral activity. Disruption of the second and fourth zinc fingers abolished ZAP's activity, whereas disruption of the first and third fingers just slightly lowered its activity.The zinc finger antiviral protein (ZAP) was originally recovered from a screen for genes conferring resistance to the infection of cells by Moloney murine leukemia virus (MLV) (11). The overexpression of ZAP rendered cells 30-fold more resistant to viral infection. An analysis to determine the step at which ZAP blocked virus infection revealed that in ZAP-expressing cells, reverse transcription and nuclear entry of the viral DNA were normal but the production of viral RNA in the cytoplasm was inhibited (11). In addition to its inhibition of MLV, ZAP potently inhibits the replication of multiple members of the Alphavirus genus of the Togaviridae family, including Sindbis virus (SIN), Semliki Forest virus, Ross River virus, and Venezuelan equine encephalitis virus (3). The expression of ZAP does not induce a broad-spectrum antiviral state, as some viruses, including herpes simplex virus type 1 and yellow fever virus, grow normally in ZAP-expressing cells (3). ZAP targets SIN at a stage after binding and penetration, and it prevents translation of the incoming viral RNA (3). Given that alphaviruses are replicated entirely in an RNA state in the cytoplasm (19) and that the production of MLV viral RNA was inhibited only in the cytoplasm, it is tempting to propose that a common mechanism occurring in the cytoplasm underlies the ZAPmediated elimination of MLV and SIN viral RNAs. However, since there is no obvious sequence homology between MLV and SIN, the common feature(s) shared by these two divergent viruses to account for their sensitivity to ZAP remained elusive.Sequence analysis revealed that in the N terminus of ZAP there are four CCCH-type zinc finger motifs. A fragment of 254 amino acids of the N terminus (NZAP) containing the four zinc finger motifs displa...

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Section: Discussionmentioning

confidence: 90%

The Zinc Finger Antiviral Protein Directly Binds to Specific Viral mRNAs through the CCCH Zinc Finger Motifs

Guo

Carroll

MacDonald

et al. 2004

J Virol

232

342

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“…These effects of single-site mutations on the RNA-binding activity of U2AF23 contrast strongly with tandemly linked CCCH-type ZFs (Michel et al 2003;Hudson et al 2004;Teplova and Patel 2008;Martinez-Lumbreras et al 2013). In the case of the TIS11d protein, no singlesite mutation on one ZF completely abolishes the RNAbinding activity of the protein overall because another ZF can still bind to the RNA molecule.…”

Section: Putative Rna-binding Sites On the U2af23 Complexmentioning

confidence: 91%

A novel 3′ splice site recognition by the two zinc fingers in the U2AF small subunit

et al. 2015

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The pre-mRNA splicing reaction of eukaryotic cells has to be carried out extremely accurately, as failure to recognize the splice sites correctly causes serious disease. The small subunit of the U2AF heterodimer is essential for the determination of 3 ′ splice sites in pre-mRNA splicing, and several single-residue mutations of the U2AF small subunit cause severe disorders such as myelodysplastic syndromes. However, the mechanism of RNA recognition is poorly understood. Here we solved the crystal structure of the U2AF small subunit (U2AF23) from fission yeast, consisting of an RNA recognition motif (RRM) domain flanked by two conserved CCCH-type zinc fingers (ZFs). The two ZFs are positioned side by side on the β sheet of the RRM domain. Further mutational analysis revealed that the ZFs bind cooperatively to the target RNA sequence, but the RRM domain acts simply as a scaffold to organize the ZFs and does not itself contact the RNA directly. This completely novel and unexpected mode of RNA-binding mechanism by the U2AF small subunit sheds light on splicing errors caused by mutations of this highly conserved protein.

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“…A TTP knockout mouse strain has a complex phenotype with features of inflammatory arthritis, both of which are A Clark Anti-inflammatory functions of glucocorticoid-induced genes Page 22 of 53 largely mediated by increased stability of TNF mRNA and increased expression of TNF protein (Carballo et al, 1998;Taylor et al, 1996). TTP recognizes the nonameric sequence UUAUUUAUU, present in the 3' untranslated region (UTR) of TNF mRNA (Brewer et al, 2004;Michel et al, 2003;Worthington et al, 2002). Many other mediators of immunity or inflammation are encoded by labile mRNAs that also have adenylate/uridylate-rich elements (AREs) in their 3' UTRs.…”

Section: Tristetraprolinmentioning

confidence: 99%

Anti-inflammatory functions of glucocorticoid-induced genes

Clark

2007

Molecular and Cellular Endocrinology

237

193

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Selective RNA Binding by a Single CCCH Zinc-Binding Domain from Nup475 (Tristetraprolin)

Cited by 59 publications

References 19 publications

The Zinc Finger Antiviral Protein Directly Binds to Specific Viral mRNAs through the CCCH Zinc Finger Motifs

The Zinc Finger Antiviral Protein Directly Binds to Specific Viral mRNAs through the CCCH Zinc Finger Motifs

A novel 3′ splice site recognition by the two zinc fingers in the U2AF small subunit

Anti-inflammatory functions of glucocorticoid-induced genes

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