Proteomic analysis of NME1/NDPK A null mouse liver: evidence for a post-translational regulation of annexin IV and EF-1Bα

Abstract

NME/NDPK family proteins are involved in the control of intracellular nucleotide homeostasis as well as in both physiological and pathological cellular processes, such as proliferation, differentiation, development, apoptosis, and metastasis dissemination, through mechanisms still largely unknown. One family member, NME1/NDPK-A, is a metastasis suppressor, yet the primary physiological functions of this protein are still missing. The purpose of this study was to identify new NME1/NDPK-A-dependent biological functions and pathways regulated by this gene in the liver. We analyzed the proteomes of wild-type and transgenic NME1-null mouse livers by combining two-dimensional gel electrophoresis and mass spectrometry (matrix-assisted laser desorption/ionization time of flight and liquid chromatography-tandem mass spectrometry). We found that the levels of three proteins, namely, phenylalanine hydroxylase, annexin IV, and elongation factor 1 Bα (EF-1Bα), were strongly reduced in the cytosolic fraction of NME1(-/-) mouse livers when compared to the wild type. This was confirmed by immunoblotting analysis. No concomitant reduction in the corresponding messenger RNAs or of total protein level was observed, however, suggesting that NME1 controls annexin IV and EF-1Bα amounts by post-translational mechanisms. NME1 deletion induced a change in the subcellular location of annexin IV in hepatocytes resulting in enrichment of this protein at the plasma membrane. We also observed a redistribution of EF-1Bα in NME1(-/-) hepatocytes to an intracytoplasmic compartment that colocalized with a marker of the reticulum endoplasmic. Finally, we found reduced expression of annexin IV coincident with decreased NME1 expression in a panel of different carcinoma cell lines. Taken together, our data suggest for the first time that NME1 might regulate the subcellular trafficking of annexin IV and EF-1Bα. The potential role of these proteins in metastatic dissemination is discussed.

Metastasis suppressor NM23 limits oxidative stress in mammals by preventing activation of stress‐activated protein kinases/JNKs through its nucleoside diphosphate kinase activity

“…Of note, livers of untreated mice of both genotypes were histologically normal as we reported previously ( Fig. 2A) (31). Although the hepatic abnormalities were modest, early mortality of NME1 mutant mice was frequent.…”

Non-translational Connections of eEF1B in the Cytoplasm and Nucleus of Cancer Cells

“…This protein interacts with nuclear/cytoplasmic prolylisomerase FKBP25 (FKBP3) (Galat et al, 2014), which is a nucleic acid binding protein involved in both repair of DNA double-strand breaks (Dilworth et al, 2019) and microtubule polymerization (Dilworth et al, 2018). Depletion of nucleoside diphosphate kinase A (NME1) lead to a redistribution of eEF1Bα from the cytoplasm to the endoplasmic reticulum fraction in mouse liver (Bruneel et al, 2011). Importantly, nucleoside diphosphate kinase A can be transferred to the nucleus where it promotes the non-homologous end joining of the DNA doublestrand breaks (Xue et al, 2019) and takes part in transcription (Puts et al, 2018).…”

“…This apparently reflects cytonucleo shuttling of subunits involved into these processes. DNA repair may implicate both eEF1Bβ and eEF1Bα (Bruneel et al, 2011;Galat et al, 2014;Kapustian et al, 2016;Kapustian et al, 2017). This suggests an involvement of the whole eEF1B complex, as these subunits interact with each other via eEF1Bγ.…”

LncRNA HOXD‐AS1 promotes oral squamous cell carcinoma by sponging miR‐203a‐5p

“…Annexin A4 is a type of calcium‐dependent phospholipid binding protein and plays an important role in membrane‐related events, including membrane permeability, exocytosis, and endocytosis (Wei et al., 2015). It has been reported that Annexin A4 is involved in the development, progression, and chemoresistance of human cancers by interacting with cancer‐related signaling pathways (Bruneel et al., 2011; Kim et al., 2009; Masuishi et al., 2011; Wei et al., 2015). Therefore, Annexin A4 is considered a potential therapeutic target for human cancers (Wei et al., 2015).…”