TINCR encodes the ubiquitin-like protein TUBL.

(A) Smoothed PhyloCSF peaks for human and mouse TINCR. (B) Comparison of the predicted amino acid sequence of human TUBL with that of the mouse, avian, reptile, and amphibian proteins. Arrowheads indicate the first and second methionines of mouse TUBL. Amino acids that differ between species are shown in red. (C) Amino acid sequence similarity for human TUBL and other human ubiquitin-like proteins compared with human ubiquitin. (D) Immunoblot (IB) analysis of HEK293T cells transiently transfected with expression vectors for mouse TINCR with a FLAG epitope tag sequence inserted at the COOH-terminus (C) of the TUBL ORF or for its ΔAUG mutants. The analysis was performed with M2 antibodies to FLAG and antibodies to HSP90 (loading control). (E) Immunofluorescence analysis of FLAG in HeLa cells expressing FLAG epitope–tagged mouse TUBL. Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI). Scale bar, 20 μm. (F) Immunoblot analysis of HEK293T cells transiently transfected with expression vectors for human (h) or mouse (m) TUBL with or without an NH₂ (N)–or COOH (C)–terminal V5 epitope tag or for human NEDD8 with an NH₂-terminal V5 tag or its mutant (ΔGG) lacking the diglycine motif. (G) Lysates of tissues from wild-type (Tubl^+/+) mice or mice harboring a FLAG epitope tag sequence at the COOH-terminus of the TUBL ORF (Tubl^FLAG/FLAG mice) were subjected to immunoprecipitation (IP) with antibodies to FLAG, and the resulting precipitates as well as the original lysates (3% of input for immunoprecipitation) were subjected to immunoblot analysis with antibodies to FLAG and to HSP70 (loading control).