A monoclonal antibody to chicken gizzard desmin that recognizes intermediate filaments and nuclear granules in BHK21/C13 cells.

Abstract

ABSTRACT.One hybridoma (AC54), which produces monoclonal antibody (MAb) that recognizes both intermediate filaments (IFs) and nuclear granules in BHK21/C13 cells, and two hybridomas (AC19 and AC36) which produce MAbs that recognize IFs only, were obtained by using a crude actin preparation from chicken gizzard as an antigen. In immunoblotting, both the AC54 and AC19 MAbs reacted with the 52 kD protein (desmin) and some other proteins in gizzard and BHK21/C13 cells. Indirect immunofluorescent microscopy of BHK21/C13 cells showed that the cytoplasmic filaments stained by these MAbs were IFs based on their colchicine-induced whorl formation. The ability of AC54 MAb to recognize IFs was more limited than that of AC19 MAb. The nuclear granules recognized by AC54 MAb were in a different location than the cytoplasmic IFs and sometimes were concentrated in the nucleolus.These results indicate that AC54 MAb is an anti-desmin MAb that reacts with some desmin-related proteins; that it recognizes IFs differently than AC19 MAb, another anti-desmin MAb; and that it recognizes nuclear granules in locations where desmin or desmin-related protein has not yet been reported.

Desmin enters the nucleus of cardiac stem cells and modulates Nkx2.5 expression by participating in transcription factor complexes that interact with thenkx2.5gene

“…Nuclear transport and presence of IF proteins has been demonstrated for tail-less cytokeratin 8, 18, and 19 in 3T3 fibroblasts ( Bader et al, 1991 ), for vimentin in nasopharyngeal carcinoma lymph node metastasis ( Luo et al, 2012 ), for nestin in various brain tumors ( Krupkova et al, 2011 ), and for desmin in BHK21 cells ( Kamei, 1986 ).…”

“…This notion is supported by data demonstrating that desmin and vimentin interact with DNA in vitro ( Tolstonog et al, 2005 ; Traub and Shoeman, 1994b ). Desmin has been detected in nuclei of BHK21 cells ( Kamei, 1986 ) and nestin in nuclei of brain tumor cells ( Krupkova et al, 2011 ). It has been suggested that vimentin enters the nucleus via a piggyback mechanism ( Hartig et al, 1998 ) and binds to DNA via its amino-terminal domain particularly at the nuclear matrix attachment regions ( Tolstonog et al, 2001 ).…”