The effect of including the C2 insert of nonmuscle myosin II-C on neuritogenesis.
The functional role of the C2 insert of nonmuscle myosin II-C (NM II-C) is poorly understood. Here, we report for the first time that the expression of the C2 insert-containing isoform, NM II-C1C2, is inducible in Neuro-2a cells during differentiation both at mRNA and protein levels. Immunoblot and RT-PCR analysis reveal that expression of NM II-C1C2 peaks between days 3 and 6 of differentiation. Localization of NM II-C1C2 in Neuro-2a cells suggests that the C2 insert-containing isoform is localized in the cytosol and along the neurites, specifically at the adherence point to substratum. Inhibition of endogenous NM II-C1C2 using siRNA decreases the neurite length by 43% compared with control cells treated with nonspecific siRNA. Time lapse image analysis reveals that neurites of C2-siRNA-treated cells have a net negative change in neurite length per minute, leading to a reduction of overall neurite length. During neuritogenesis, NM II-C1C2 can interact and colocalize with β1-integrin in neurites. Altogether, these studies indicate that NM II-C1C2 may be involved in stabilizing neurites by maintaining their structure at adhesion sites.
Department of Biological Chemistry, Indian Association for the Cultivation of Science, Jadavpur, Kolkata-32, India.
Shekhar
S
Saha
Department of Biological Chemistry, Indian Association for the Cultivation of Science, Jadavpur, Kolkata-32, India.
Sumit K
SK
Dey
Infectious Diseases and Immunology Division, Indian Institute of Chemical Biology, Kolkata-32, India.
Arunima
A
Biswas
Department of Biological Chemistry, Indian Association for the Cultivation of Science, Jadavpur, Kolkata-32, India.
Provas
P
Das
Department of Biological Chemistry, Indian Association for the Cultivation of Science, Jadavpur, Kolkata-32, India.
Mahua R
MR
Das
Department of Biological Chemistry, Indian Association for the Cultivation of Science, Jadavpur, Kolkata-32, India. Electronic address: bcssj@iacs.res.in.
Siddhartha S
SS
Jana
D017398:Alternative Splicing; D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods; D008954:Models, Biological; D018995:Myosin Heavy Chains / Q000737:chemistry* / Q000378:metabolism; D024462:Myosin Type II / Q000737:chemistry* / Q000378:metabolism; D009218:Myosins / Q000378:metabolism; D016501:Neurites / Q000378:metabolism; D009474:Neurons / Q000378:metabolism; D020033:Protein Isoforms; D011554:Pseudopodia / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D014162:Transfection
C483905:Myh14 protein, mouse; D020033:Protein Isoforms; D034741:RNA, Small Interfering; D024462:Myosin Type II; D018995:Myosin Heavy Chains; D009218:Myosins