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Murine isoforms of UDP-GlcNAc 2-epimerase/ManNAc kinase: Secondary structures, expression profiles, and response to ManNAc therapy

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Abstract

The bifunctional enzyme UDP-GlcNAc 2-epimerase/ManNAc kinase (GNE) catalyzes the first two committed steps in sialic acid synthesis. Non-allosteric GNE gene mutations cause the muscular disorder GNE myopathy (also known as hereditary inclusion body myopathy), whose exact pathology remains unknown. Increased knowledge of GNE regulation, including isoform regulation, may help elucidate the pathology of GNE myopathy. While eight mRNA transcripts encoding human GNE isoforms are described, we only identified two mouse Gne mRNA transcripts, encoding mGne1 and mGne2, homologous to human hGNE1 and hGNE2. Orthologs of the other human isoforms were not identified in mice. mGne1 appeared as the ubiquitously expressed, major mouse isoform. The mGne2 encoding transcript is differentially expressed and may act as a tissue-specific regulator of sialylation. mGne2 expression appeared significantly increased the first 2 days of life, possibly reflecting the high sialic acid demand during this period. Tissues of the knock-in Gne p.M712T mouse model had similar mGne transcript expression levels among genotypes, indicating no effect of the mutation on mRNA expression. However, upon treatment of these mice with N-acetylmannosamine (ManNAc, a Gne substrate, sialic acid precursor, and proposed therapy for GNE myopathy), Gne transcript expression, in particular mGne2, increased significantly, likely resulting in increased Gne enzymatic activities. This dual effect of ManNAc supplementation (increased flux through the sialic acid pathway and increased Gne activity) needs to be considered when treating GNE myopathy patients with ManNAc. In addition, the existence and expression of GNE isoforms needs consideration when designing other therapeutic strategies for GNE myopathy.

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Abbreviations

A. avenae :

Acidovorax avenae

B2M:

β2 microglobulin

BLAST:

Basic Local Alignment Search Tool

B. subtilis :

Bacillus subtilis

CDM:

Consensus Data Mining

CMP:

Cytidine monophosphate

DMRV:

Distal Myopathy with Rimmed Vacuoles

DYRK2:

Dual-specificity tyrosine phosphorylation regulated kinase 2

E:

Embryonic day

E. coli :

Escherichia coli

FDM:

Fragment Database Mining

GCNA:

N-acetyl-β-D-glucosaminidase

GlcNAc:

N-acetylglucosamine

GNE:

UDP-GlcNAc 2-epimerase/ManNAc kinase

GOR:

Garnier-Osguthorpe-Robson

H. arsenicoxydans :

Herminiimonas arsenicoxydans

HIBM:

Hereditary Inclusion Body Myopathy

His:

Histidine

H. sapiens :

Homo sapiens

Min:

Minutes

ManNAc:

N-acetylmannosamine

NCBI:

National Center for Biotechnology Information

Neu5Ac:

N-acetylneuraminic acid

OMIM:

Online Mendelian Inheritance in Man

P:

Postnatal day

P. carotovorum :

Pectobacterium carotovorum

PCR:

Polymerase chain reaction

Pdb:

Protein databank

PRPP:

Phosphoribosylpyrophosphate

PSIPRED:

Protein Structure Initiative Prediction

qPCR:

Quantitative real-time polymerase chain reaction

RNA:

Ribonucleic acid

SGK1:

Serum and glucocorticoid-induced kinase 1

S. gordonii :

Streptococcus gordonii

T. maritime :

Thermotoga maritima

UDP:

Uridine diphosphate

VAV1:

Vav-family member of guanine nucleotide exchange factors

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Acknowledgments

We greatly appreciate the expert laboratory work of Dr. Shelley Hoogstraten-Miller, Katherine Berger, Katherine Patzel, and Adrian Astiz-Martinez. The authors thank Theresa Calhoun for her skilled assistance with mouse maintenance. We thank the HIBM Research group (Encino, CA) for providing the knock-in Gne p.M712T mouse model. This work was performed in partial fulfillment of the requirements for a PhD degree of T.Y., Sackler Faculty of Medicine, Tel Aviv University, Israel. This study was supported by the Intramural Research Program of the National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland, United States (T.Y., K.J., T.K.N., C.C., W.A.G., and M.H.) and Research Funds of The School of Theoretical Modeling, Chevy Chase, Maryland, United States (N.K.).

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Authors and Affiliations

  1. Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, 10 Center Drive, MSC 1851 Bld 10, Rm 10C103, Bethesda, MD, 20892-1851, USA

    Tal Yardeni, Katherine Jacobs, Terren K. Niethamer, Carla Ciccone, William A. Gahl & Marjan Huizing

  2. Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, 69978, Israel

    Tal Yardeni & Yair Anikster

  3. Department of Biophysics, The School of Theoretical Modeling, Chevy Chase, MD, 20825, USA

    Natalya Kurochkina

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  1. Tal Yardeni
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Correspondence to Marjan Huizing.

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Yardeni, T., Jacobs, K., Niethamer, T.K. et al. Murine isoforms of UDP-GlcNAc 2-epimerase/ManNAc kinase: Secondary structures, expression profiles, and response to ManNAc therapy. Glycoconj J 30, 609–618 (2013). https://doi.org/10.1007/s10719-012-9459-1

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