Microinjection of specific anti-IMPDH2 antibodies induces disassembly of cytoplasmic rods/rings that are primarily stationary and stable structures

Microinjection of specific anti-IMPDH2 antibodies induces disassembly of cytoplasmic rods/rings that are primarily stationary and stable structures

Author Keppeke, Gerson Dierley Autor UNIFESP Google Scholar
Andrade, Luis Eduardo C. Autor UNIFESP Google Scholar
Grieshaber, Scott S. Google Scholar
Chan, Edward K. L. Google Scholar
Institution Univ Florida
Universidade Federal de São Paulo (UNIFESP)
Fleury Med & Hlth Labs
Univ Idaho
Abstract Background: Our laboratory previously reported interesting rods 3-10 mu m long and rings 2-5 mu m diameter (RR) in the cytoplasm of mammalian cells. Experimental evidence show that both inosine-5'-monophosphate dehydrogenase 2 (IMPDH2) and cytidine triphosphate synthetase (CTPS) are components of RR structures. Several cell types, including mouse embryonic stem cells, and cell lines, such as mouse 3 T3 and rat NRK, naturally present RR structures, while other cells can present RR when treated with compounds interfering with GTP/CTP biosynthetic pathways. in this study, we aimed to investigate the dynamic behavior of these RR in live cells.Results: RR were detected in > 90% of COS-7 and HeLa cells treated with 1 mM ribavirin or 6-Diazo-5-oxo-L-norleucine (DON) for 24 h, and in 75% of COS-7 cells treated with 1 mM mycophenolic acid (MPA) for the same period of time. Microinjection of affinity-purified anti-IMPDH2 antibodies in live COS-7 cells treated with ribavirin, DON, or MPA showed mature forms of RR presented as stable and stationary structures in 71% of cells. in the remaining 29% of cells, RR acquired erratic movement and progressively disassembled into fragments and disappeared within 10 min. the specific stationary state and antibody-dependent disassembling of RR structures was independently confirmed in COS-7 and HeLa cells transfected with GFP-tagged IMPDH2.Conclusions: This is the first demonstration of disassembly of RR structures upon microinjection of anti-IMPDH2 antibodies that led to the disappearance of the molecular aggregates. the disassembly of RR after microinjection of anti-IMPDH2 antibody further strengthens the notion that IMPDH2 are major building blocks of RR. Using two independent methods, this study demonstrated that the induced RR are primarily stationary structures in live cells and that IMPDH2 is a key component of RR.
Keywords CTP synthase
Inosine monophosphate dehydrogenase
Intracellular compartment
Mycophenolic acid
Language English
Sponsor Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Grant number CAPES: 9028-11-0
FAPESP: 2011/12448-0
Date 2015-01-05
Published in Cell and Bioscience. London: Biomed Central Ltd, v. 5, 11 p., 2015.
ISSN 2045-3701 (Sherpa/Romeo, impact factor)
Publisher Biomed Central Ltd
Extent 11
Origin http://dx.doi.org/10.1186/2045-3701-5-1
Access rights Open access Open Access
Type Article
Web of Science ID WOS:000349257100001
URI http://repositorio.unifesp.br/handle/11600/38630

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