Structure of the carboxy-terminal LIM domain from the cysteine rich protein CRP

Abstract

The three dimensional solution structure of the carboxy terminal LIM domain of the avian Cysteine Rich Protein (CRP) has been determined by nuclear magnetic resonance spectroscopy. The domain contains two zinc atoms bound independently in CCHC (C = Cys, H = His) and CCCC modules. Both modules contain two orthogonally-arranged antiparallel beta-sheets, and the CCCC module contains an alpha-helix at its C terminus. The modules pack due to hydrophobic interactions forming a novel global fold. The structure of the C-terminal CCCC module is essentially identical to that observed for the DNA-interactive CCCC modules of the GATA-1 and steroid hormone receptor DNA binding domains, raising the possibility that the LIM motif may have a DNA binding function.

Inhibition of activated Ras-induced neuronal differentiation of PC12 cells by the LIM domain of LIM-kinase 1

“…The LIM motif was de®ned as a structural motif composed of two adjacent zinc ®ngers separated by a two-amino-acids linker and with a consensus sequence (CX 2 CX 16 ± 23 HX 2 C)X 2 (CX 2 CX 16 ± 21 -CX 2 ± 3 C/H/D) (Sa nchez-Garcia and Rabbitts, 1994; Dawid et al, 1995). Nuclear magnetic resonance analysis revealed the three-dimensional structure of the LIM motif; it is constructed of antiparallel b sheets with two zinc ion-binding modules (Pe rez-Alvarado et al, 1994). The LIM motif was found in diverse proteins, including homeodomain-containing transcription factors, cytoskeletal proteins, and other signaling proteins, which were found to be involved in cell fate determination, growth regulation, and oncogenesis (Sa nchezGarcia and Rabbitts, 1994; Dawid et al, 1995).…”

Specificity of single LIM motifs in targeting and LIM/LIM interactions in situ.

“…This is nearly identical to the LIM motif, except for the spacer region between the two zinc fingers, which consists of only two residues in the LIM motif. Solution nuclear magnetic resonance (NMR) analysis revealed that the second finger of GATA-1 folds into a structure that is essentially identical to that of the second finger in C2 of CRP (Perez-Alvarado et al 1994). …”

A novel IL-10 signalling mechanism regulates TIMP-1 expression in human prostate tumour cells

“…The LIM domain has been found to mediate 'protein -protein' interactions of cytoplasmic and nuclear proteins. NMR analysis of the solution structure of the c-terminal LIM domain of two of these proteins, CRP1 (Perez-Alvarado et al, 1994) and CRP2 (Konrat et al, 1997;Weiskirchen et al, 1997;Wadman et al, 1994), has shown that two zinc-fingers constitute a LIM domain, which represent two separated structural entities held together in a distinct configuration by hydrophobic interactions (Dawid et al, 1998). Moreover, the c-terminal LIM domain in CRP1 and CRP2 has been found to closely resemble the zinc-finger in the DNA-binding domain of GATA1, suggesting that the LIM domain might specifically bind DNA (Weiskirchen et al, 1995).…”

“…Several studies have shown that LIM-homeodomain proteins can activate the expression of specific genes (i.e. insulin) or direct cellular differentiation in cooperation with other nuclear factors (Perez-Alvarado et al, 1994). Other examples include, E47/Pan-1 and Lmx1.1 transcription factors, which have been shown to interact with LMO2, a LIM only protein, and with TAL1, LYL1, and LYL2 to regulate gene activation (Sadler et al, 1992;Warren et al, 1994;Larson et al, 1996;Johnson et al, 1997;Dawid et al, 1998).…”