RLIM inhibits functional activity of LIM homeodomain transcription factors via recruitment of the histone deacetylase complex

Abstract

LIM domains1 are required for both inhibitory effects on LIM homeodomain transcription factors and synergistic transcriptional activation events1,2,3,4. The inhibitory actions of the LIM domain can often be overcome by the LIM co-regulator known as CLIM2, LDB1 and NLI (referred to hereafter as CLIM2; refs 2, 3, 4). The association of the CLIM cofactors with LIM domains does not, however, improve the DNA-binding ability of LIM homeodomain proteins4,5, suggesting the action of a LIM-associated inhibitor factor. Here we present evidence that LIM domains are capable of binding a novel RING-H2 zinc-finger protein, Rlim (for RING finger LIM domain-binding protein), which acts as a negative co-regulator via the recruitment of the Sin3A/histone deacetylase corepressor complex. A corepressor function of RLIM is also suggested by in vivo studies of chick wing development. Overexpression of the gene Rnf12, encoding Rlim, results in phenotypes similar to those observed after inhibition of the LIM homeodomain factor LHX2, which is required for the formation of distal structures along the proximodistal axis, or by overexpression of dominant-negative CLIM1. We conclude that Rlim is a novel corepressor that recruits histone deacetylase-containing complexes to the LIM domain.

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Figure 1: Rlim, a novel LIM domain-binding RING zinc-finger protein.
Figure 2: Comparison of Rnf12 (encoding Rlim), Ldb2 (encoding Clim1) and Ldb1 (encoding Clim2) expression patterns during mouse embryogenesis by in situ hybridizations on serial sections at E7.5 (a) and E15.5 (bg).
Figure 3: Rlim repressor function.
Figure 4: RNF12 regulates chick limb development.
Figure 5: Overexpression of dominant-negative CLIM1 or LHX2 perturbs limb outgrowth.

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Acknowledgements

We thank C. Kintner for initial X. laevis mRNA injections; O. Bernard for the Kiz-1 expression plasmid; C. Nelson for cell culture; P. Meyers for assistance in figure preparation; M. Fisher for assistance in manuscript preparation; E. De Robertis, E. Osmundson, M. Ruchhoef, S. O'Connell, M. Wegner and R. Stan for providing reagents and advice; and A. Ryan and H. Ostendorff for comments on the manuscript. C.C. was a fellow of the LALOR Foundation. This work is supported by NIH grants to B.A. (AR044882 and AR02080), J.C.I.B. and M.G.R.; by the Irving Weinstein Foundation to B.A.; and by a G. Harold and L.Y. Mathers Charitable Foundation grant to J.C.I.B.

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Correspondence to Ingolf Bach or Juan Carlos Izpisúa Belmonte or Michael G. Rosenfeld.

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Bach, I., Rodriguez-Esteban, C., Carrière, C. et al. RLIM inhibits functional activity of LIM homeodomain transcription factors via recruitment of the histone deacetylase complex. Nat Genet 22, 394–399 (1999). https://doi.org/10.1038/11970

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