Isolation, molecular characterization, and tissue-specific expression of ECP-51 and ECP-54 (TIP49), two homologous, interacting erythroid cytosolic proteins

doi:10.1016/S0167-4781(99)00104-9

Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression

Volume 1446, Issue 3, 3 September 1999, Pages 365-370

https://doi.org/10.1016/S0167-4781(99)00104-9 Get rights and content

Abstract

We isolated two proteins, ECP-51 and ECP-54, from human erythrocyte cytosol by affinity chromatography using a peptide of the integral membrane protein stomatin as bait. Partial amino acid sequence information obtained by microsequencing allowed us to clone the respective cDNAs. Analysis of the nucleotide sequences revealed that ECP-51 and ECP-54 are homologous (44.2% amino acid identity) and contain ATP-binding sites. ECP-54 was identified as TIP49/RUVBL1/NMP238, which is a component of a large nuclear protein complex, possibly the RNA polymerase II holoenzyme; ECP-51 is a novel protein. Using the two-hybrid system, we showed that these proteins interact with each other. The interaction of ECP-51 and ECP-54 with the stomatin peptide and the localization to the nucleus and cytoplasm suggest an additional function for these proteins as chaperone components.

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While in another report human RuvBL1/TIP49a was present not only in the nucleus, as expected, but was also concentrated at the centrosome (Gartner et al., 2003). Some other studies also reported its partial (Boulon et al., 2008; Sigala et al., 2005) extra nuclear localization (Salzer et al., 1999). Thus on the basis of RuvB1 localization in P. falciparum it can be expected that it may be involved in the nuclear function.
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Through a comparative study of the proteome of HCC with that of the peri-tumour liver, we identified the deregulation of a number of proteins [2], especially the overexpression of RuvBL2/Reptin [3]. Reptin is also known as TIP49b [4], TIP48 [5], Reptin52 [6], Rvb2 [7], TAP54β [8], and ECP-51 [9]. It belongs to the AAA + family of ATPases (reviewed in [10,11]) and shows a limited homology to the bacterial RuvB ATP-dependent DNA helicase.
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In vivo Reptin depletion leads to tumour growth arrest. Reptin may prove a valuable target in HCC.
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Citation Excerpt :
RVB1 and RVB2 are ATP-binding proteins that belong to the AAA+ (ATPase associated with diverse cellular activities) family of ATPases. RVBs (RVB1 and RVB2) were discovered independently in multiple organisms (Bauer et al., 1998, 2000; Kanemaki et al., 1997, 1999; Qiu et al., 1998; Salzer et al., 1999; Wood et al., 2000) and implicated in many cellular pathways (Figure 1). The structure of the RVB1/RVB2 complex that has recently been elucidated suggests that RVBs could act as a scaffolding protein, explaining its appearance in diverse cellular protein complexes (Matias et al., 2006; Puri et al., 2007; Torreira et al., 2008).
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Organ size is precisely regulated during development, but the control mechanisms remain obscure. We have isolated a mutation in zebrafish, liebeskummer (lik), which causes development of hyperplastic embryonic hearts. lik encodes Reptin, a component of a DNA-stimulated ATPase complex. The mutation activates ATPase activity of Reptin complexes and causes a cell-autonomous proliferation of cardiomyocytes to begin well after progenitors have fashioned the primitive heart tube. With regard to heart growth, β-catenin and Pontin, a DNA-stimulated ATPase that is often part of complexes with Reptin, are in the same genetic pathways. Pontin reduction phenocopies the cardiac hyperplasia of the lik mutation. Thus, the Reptin/Pontin ratio serves to regulate heart growth during development, at least in part via the β-catenin pathway.

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¹: The nucleotide sequence data reported in this paper will appear in the DDBJ, EMBL and GenBank databases with accession numbers Y18417 (ECP-51) and Y18418 (ECP-54).

²: These authors contributed equally to this study.

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Short sequence-paperIsolation, molecular characterization, and tissue-specific expression of ECP-51 and ECP-54 (TIP49), two homologous, interacting erythroid cytosolic proteins1

Abstract

Biochim. Biophys. Acta

J. Biol. Chem.

Blood

Biochim. Biophys. Acta

J. Biol. Chem.

Biochim. Biophys. Acta

Blood

J. Mol. Biol.

Biochem. Biophys. Res. Commun.

Biochem. Biophys. Res. Commun.

J. Biol. Chem.

Biochem. Biophys. Res. Commun.

Methods Enzymol.

Trends Biochem. Sci.

Exp. Cell Res.

J. Biol. Chem.

J. Biol. Chem.

Short sequence-paper
Isolation, molecular characterization, and tissue-specific expression of ECP-51 and ECP-54 (TIP49), two homologous, interacting erythroid cytosolic proteins¹