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

Cell & Bioscience

Edward K L Chan¹  Scott S Grieshaber²  Luís Eduardo C Andrade³  Gerson Dierley Keppeke⁴ 

[1]Department of Oral Biology, University of Florida, 1395 Center Drive, Gainesville, FL 32610-0424, USA

[2]Department of Biological Sciences, University of Idaho, 875 Center Drive, Moscow, ID 83844, USA

[3]Immunology Division, Fleury Medicine and Health Laboratories, Avenida Gal Waldomiro Lima 508, São Paulo, SP 04102-050, Brazil

[4]Rheumatology Division, Universidade Federal de São Paulo, Rua Botucatu 740, São Paulo, SP 04023-062, Brazil

关键词: Ribavirin;    Mycophenolic acid;    Intracellular compartment;    Inosine monophosphate dehydrogenase;    Cytoophidium;    CTP synthase;   

Others  :  1135257
DOI  :  10.1186/2045-3701-5-1

 received in 2014-07-25, accepted in 2014-12-18,  发布年份 2015

【 摘 要 】

Background

Our laboratory previously reported interesting rods 3–10 μm long and rings 2–5 μm 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.

【 授权许可】

   
2015 Keppeke et al.; licensee BioMed Central.

【 图 表 】

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