Journal of Cell Science, Vol 101, Issue 4 773-784, Copyright © 1992 by Company of Biologists

JOURNAL ARTICLES

A monoclonal antibody recognizing nuclear matrix-associated nuclear bodies

N Stuurman, A de Graaf, A Floore, A Josso, B Humbel, L de Jong and R van Driel
E. C. Slater Institute for Biochemical Research, University of Amsterdam, The Netherlands.

We have isolated a monoclonal antibody, 5E10, that labels discrete spots in the interphase nucleus. By immunoblotting mAb 5E10 recognized predominantly a 126 kDa polypeptide with an isoelectric point of 5.5. Indirect immunofluorescence showed that mAb 5E10 labeled spots in many cell lines and tissues from rat or human origin, but not in cells from mouse, chicken, African green monkey, or the lower eukaryotes Saccharomyces and Dictyostelium. In the human bladder carcinoma cell line T24 the number of nuclear spots were found to be 21 +/- 10 (n = 132). In many cells spots were found also in the cytoplasm. In a small fraction of T24 cells the mAb revealed thread-like structures in addition to spots. Throughout mitosis the antigen was found to be clustered in the cytoplasm, not associated with metaphase chromosomes. The spherical structures that contain the antigen were tightly bound to the nuclear matrix. Immunogold labeling with mAb 5E10 showed that the antigen is localized in 0.3 microns diameter spherical, electron-dense structures, reminiscent of nuclear bodies. Double-labeling experiments showed that these spots do not colocalize with U1 snRNPs and centromeres. The spots did colocalize with nuclear speckles recognized by a primary biliary cirrhosis autoimmune serum, which is thought to recognize nuclear bodies. On the basis of these observations we conclude that mAb 5E10 recognizes discrete nuclear substructures, most likely nuclear bodies.
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				M. H. M. Koken, A. Reid, F. Quignon, M. K. Chelbi-Alix, J. M. Davies, J. H. S. Kabarowski, J. Zhu, S. Dong, S.-J. Chen, Z. Chen, et al. Leukemia-associated retinoic acid receptor alpha  fusion partners, PML and PLZF, heterodimerize and colocalize to nuclear bodies PNAS, September 16, 1997; 94(19): 10255 - 10260. [Abstract] [Full Text] [PDF]

				C. Gongora, G. David, L. Pintard, C. Tissot, T. D. Hua, A. Dejean, and N. Mechti Molecular Cloning of a New Interferon-induced PML Nuclear Body-associated Protein J. Biol. Chem., August 1, 1997; 272(31): 19457 - 19463. [Abstract] [Full Text] [PDF]

				A. M. Ishov, R. M. Stenberg, and G. G. Maul Human Cytomegalovirus Immediate Early Interaction with Host Nuclear Structures: Definition of an Immediate Transcript Environment J. Cell Biol., July 14, 1997; 138(1): 5 - 16. [Abstract] [Full Text] [PDF]

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				Y. Shimono, H. Murakami, Y. Hasegawa, and M. Takahashi RET Finger Protein Is a Transcriptional Repressor and Interacts with Enhancer of Polycomb That Has Dual Transcriptional Functions J. Biol. Chem., December 8, 2000; 275(50): 39411 - 39419. [Abstract] [Full Text] [PDF]

				W. T. Tse, J. Tang, O. Jin, C. Korsgren, K. M. John, A. L. Kung, B. Gwynn, L. L. Peters, and S. E. Lux A New Spectrin, beta IV, Has a Major Truncated Isoform That Associates with Promyelocytic Leukemia Protein Nuclear Bodies and the Nuclear Matrix J. Biol. Chem., June 22, 2001; 276(26): 23974 - 23985. [Abstract] [Full Text] [PDF]