Cloning and functional characterization of resistin-like molecule γ

doi:10.1016/j.bbrc.2003.12.100

Biochemical and Biophysical Research Communications

Volume 314, Issue 2, 6 February 2004, Pages 356-362

https://doi.org/10.1016/j.bbrc.2003.12.100 Get rights and content

Abstract

Resistin, a recently discovered hormone that may play a crucial role in obesity-associated diabetes, is the founding member of a novel family of cysteine-rich proteins that are secreted by specific cell types. Three other members of this family have been described to date and were termed resistin-like molecules (RELMs). Here we describe the cloning and functional characterization of RELMγ. The mouse RELMγ-cDNA encodes a protein of 117 amino acids that contains a signal peptide leading to secretion of the protein. By Northern blotting the RELMγ-mRNA is detectable in bone marrow, spleen, and lung as well as in peripheral blood granulocytes. Promyelocytic HL60 cells transfected with a RELMγ expression plasmid have an increased proliferation rate compared to mock-transfected cells and display an altered response to retinoic acid-induced granulocytic differentiation. Taken together, these data provide the first experimental evidence that RELMγ is a secreted molecule with a restricted expression pattern that may play a role in promyelocytic differentiation.

Section snippets

Materials and methods

Cloning of the RELMγ-cDNA. Total RNA was extracted from femora of three-day-old C57Bl/6 mice using the TRIzol reagent (Invitrogen). This RNA was transcribed into cDNA by reverse transcription and used as a template in a subsequent PCR. The primers used for this reaction were 5^′-TTGTTTCCTTTTCATCTTCGTCTC-3^′ and 5^′-CAGTGGCAAGTATTTCCATTCCG-3^′ and corresponded to the most conserved regions of the mouse cDNA sequences of resistin, RELMα, and RELMβ. PCR conditions were as follows: 94 °C for 30 s; 50 °C

Cloning of the RELMγ-cDNA

Based on the clinical observation that there is an association between type 2 diabetes and increased bone mineral density [12], our initial hypothesis was that bone cells could produce a cell-specific resistin-like molecule. To analyze this possibility we isolated cDNA from femora of newborn mice and performed a PCR using primers corresponding to the most conserved regions of the mouse cDNAs encoding resistin, RELMα, and RELMβ. The resulting PCR product was inserted into the cloning vector pCR

Discussion

Since the discovery of resistin as a potential insulin-antagonizing hormone [1], the resistin protein family has gained a lot of attention in the scientific community. The fact that the members of this family are secreted molecules expressed in a tissue-specific manner raised the hypothesis that they may serve important functions as signaling molecules [3], [4]. Here, we describe the cloning and functional characterization of murine RELMγ. Like the other members of the family, RELMγ is a

Acknowledgements

This work was supported by a grant from the Erich and Gertrud Roggenbuck Foundation.

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RELM genes encode secreted proteins of 105–138 amino acids in size with 3 main domains: an amino (N) terminal signal sequence, a variable middle section, and a conserved carboxyl (C) terminal. The C terminal is comprised of a cysteine signature motif sequence shared by all RELM family members (C-X11-C-X8-C-X-C-X3-C-X10-C-X-C-X-C-X9-CC-X3–6-END), which is proposed to be critical for disulfide bond formation and protein folding [3,6,7]. The crystal structures of mouse resistin and RELMβ have been solved, revealing that they form trimers linked together via disulfide bonds to form hexameric assemblies [8].
The Resistin-Like Molecules (RELM) α, β, and γ and their namesake, resistin, share structural and sequence homology but exhibit significant diversity in expression and function within their mammalian host. RELM proteins are expressed in a wide range of diseases, such as: microbial infections (eg. bacterial and helminth), inflammatory diseases (eg. asthma, fibrosis) and metabolic disorders (eg. diabetes). While the expression pattern and molecular regulation of RELM proteins are well characterized, much controversy remains over their proposed functions, with evidence of host-protective and pathogenic roles. Moreover, the receptors for RELM proteins are unclear, although three receptors for resistin, decorin, adenylyl cyclase-associated protein 1 (CAP1), and Toll-like Receptor 4 (TLR4) have recently been proposed. In this review, we will first summarize the molecular regulation of the RELM gene family, including transcription regulation and tissue expression in humans and mouse disease models. Second, we will outline the function and receptor-mediated signaling associated with RELM proteins. Finally, we will discuss recent studies suggesting that, despite early misconceptions that these proteins are pathogenic, RELM proteins have a more nuanced and potentially beneficial role for the host in certain disease settings.
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Grem2 promotes the apoptosis of medial smooth muscle cells, and its enhancement of gene expression may be explained by this mechanism (Maciel et al., 2009). It was also suggested that bone marrow-derived vascular progenitor cells are involved in the pathogenesis of atherosclerosis (Sata, 2003), raising the possibility that the differentiation-inducing activity of promyelocytes with RELM-γ is also involved (Schinke et al., 2004). Systolic blood pressure in HFDS-fed SHHR did not differ significantly from that in control SHHR.
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¹: These authors contributed equally to this work.

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Cloning and functional characterization of resistin-like molecule γ

Abstract

Section snippets

Materials and methods

Cloning of the RELMγ-cDNA

Discussion

Acknowledgements

Genomics

J. Biol. Chem.

J. Biol. Chem.

Blood

Leuk. Res.

Blood

The hormone resistin links obesity to diabetes

Nature

Resistin: molecular history and prognosis

J. Mol. Med.

FIZZ1, a novel cysteine-rich secreted protein associated with pulmonary inflammation, defines a new gene family

EMBO J.

A family of tissue-specific resistin-like molecules

Proc. Natl. Acad. Sci. USA