Expression analysis of ATAD3 isoforms in rodent and human cell lines and tissues.
ATAD3 (ATPase family AAA-Domain containing protein 3) is a mitochondrial inner membrane ATPase with unknown but vital functions. Initial researches have focused essentially on the major p66-ATAD3 isoform, but other proteins and mRNAs are described in the data banks. Using a set of anti-peptide antibodies and by the use of rodent and human cell lines and organs, we tried to detail ATAD3 gene expression profiles and to verify the existence of the various ATAD3 isoforms. In rodent, the single ATAD3 gene is expressed as a major isoform of 67 kDa, (ATAD3l; long), in all cells and organs studied. A second isoform, p57-ATAD3s (small), is expressed specifically throughout brain development and in adult, and overexpressed around the peri-natal period. p57-ATAD3s is also expressed in neuronal and glial rodent cell lines, and during in vitro differentiation of primary cultured rat oligodendrocytes. Other smaller isoforms were also detected in a tissue-specific manner. In human and primates, ATAD3 paralogues are encoded by three genes (ATAD3A, 3B and 3C), each of them presenting several putative variants. Analyzing the expression of ATAD3A and ATAD3B with four specific anti-peptide antibodies, and comparing their expressions with in vitro expressed ATAD3 cDNAs, we were able to observe and define five isoforms. In particular, the previously described p72-ATAD3B is confirmed to be in certain cases a phosphorylated form of ATAD3As. Moreover, we observed that the ATAD3As phosphorylation level is regulated by insulin and serum. Finally, exploring ATAD3 mRNA expression, we confirmed the existence of an alternative splicing in rodent and of several mRNA isoforms in human. Considering these observations, we propose the development of a uniform denomination for ATAD3 isoforms in rodent and human.
INSERM U1055, Laboratoire de Bioénergétique Fondamentale et Appliquée (LBFA) et SFR Biologie Environnementale et Systémique (BEeSy), Joseph Fourier University, Grenoble F-38041, France.
Shuijie
S
Li
INSERM U1055, Laboratoire de Bioénergétique Fondamentale et Appliquée (LBFA) et SFR Biologie Environnementale et Systémique (BEeSy), Joseph Fourier University, Grenoble F-38041, France; Grenoble University Hospital, Grenoble F-38043, France.
Fredéric
F
Lamarche
Département de Microbiologie et Infectiologie, Faculté de Médecine, Université de Sherbrooke, Sherbrooke, QC J1J3P9, Canada.
Romain
R
Charton
Grenoble Institut des Neurosciences (GIN), INSERM U836-UJF-CEA-CHU, Université Joseph Fourier, 38706 La Tronche Cedex, France.
Christian
C
Delphin
Ludwig-Maximilians-Universität München, ENT Research, ENT Clinics, Marchioninistr. 15, D-81377 München, Germany.
Olivier
O
Gires
Department of Cell and Developmental Biology, University of Wisconsin, Aurora, CO 80045, USA.
Arnaud
A
Hubstenberger
INSERM U1055, Laboratoire de Bioénergétique Fondamentale et Appliquée (LBFA) et SFR Biologie Environnementale et Systémique (BEeSy), Joseph Fourier University, Grenoble F-38041, France.
Uwe
U
Schlattner
INSERM U1055, Laboratoire de Bioénergétique Fondamentale et Appliquée (LBFA) et SFR Biologie Environnementale et Systémique (BEeSy), Joseph Fourier University, Grenoble F-38041, France. Electronic address: Denis.Rousseau@ujf-grenoble.fr.
Denis
D
Rousseau
D000074183:ATPases Associated with Diverse Cellular Activities; D000251:Adenosine Triphosphatases / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology; D002460:Cell Line; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D051379:Mice; D024101:Mitochondrial Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D008969:Molecular Sequence Data; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D017386:Sequence Homology, Amino Acid
C549686:ATAD3A protein, human; C519737:ATAD3B protein, human; D000906:Antibodies; C582415:Atad3a protein, mouse; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D020033:Protein Isoforms; D012333:RNA, Messenger; D000251:Adenosine Triphosphatases; D000074183:ATPases Associated with Diverse Cellular Activities
2′, 3′-cyclic nucleotide 3′-phosphodiesterase; AAA-ATPase; ATAD3; ATPase family AAA-Domain containing protein 3; CNPase; E3PD; Exon 3-encoded Protein Domain; GAPDH; Glyceraldehyde-3-phosphate dehydrogenase; Mitochondria; OPCs; ORF; Oligodendrocyte Progenitor Cells; Open Reading Frame; PKC; Protein Kinase C; REF; RT-PCR; Rat Embryo Fibroblast; Reverse Transcription-Polymerase Chain Reaction; S100