Transcription-dependent redistribution of nuclear protein 4.1 to SC35-enriched nuclear domains.
Protein 4.1, originally identified as a component of the membrane-skeleton of the red blood cell, has also been localized in the nucleus of mammalian cells. To learn more about nuclear 4.1 protein, we have analyzed the nature of its association with the nuclear structure in comparison with SC35 and snRNP antigens, splicing proteins of the nuclear speckle domains. When MDCK or HeLa cells were digested with DNase I and washed in the presence of high salt (2 M NaCl), snRNP antigens were extracted whereas protein 4.1 and SC35 remained colocalizing in nuclear speckles. In cells treated with RNase A or heat shocked, nuclear 4.1 distribution also resembled that of SC35. Experiments carried out in transcriptionally active nuclei showed that protein 4.1 distributed in irregularly shaped speckles which appeared to be interconnected. During transcriptional inhibition, protein 4.1 accumulated in rounded speckles lacking interconnections. When cells were released from transcriptional inhibition, protein 4.1 redistributed back to the interconnected speckle pattern of transcriptionally active cells, as it was also observed for SC35. Finally, coprecipitation of 4.1 and SC35 proteins from RNase A digested HeLa nuclei further indicates that these two proteins are associated, forming part of the nuclear speckle domains to which they attach more tightly than snRNP antigens.
Lallena
M J
MJ
Centro de Biología Molecular Severo Ochoa (CSIC-UAM), Universidad Autónoma de Madrid, Spain.
Correas
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D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D003598:Cytoskeletal Proteins; D003850:Deoxyribonuclease I / Q000378:metabolism; D004285:Dogs; D019084:Fluorescent Antibody Technique, Indirect; D006367:HeLa Cells; D018869:Heat-Shock Response; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*; D009479:Neuropeptides; D009687:Nuclear Proteins / Q000378:metabolism*; D011233:Precipitin Tests; D012259:Ribonuclease, Pancreatic / Q000378:metabolism; D012261:Ribonucleoproteins; D017411:Ribonucleoproteins, Small Nuclear / Q000378:metabolism*; D000068103:Serine-Arginine Splicing Factors; D013696:Temperature; D014158:Transcription, Genetic
D003598:Cytoskeletal Proteins; D008565:Membrane Proteins; D009479:Neuropeptides; D009687:Nuclear Proteins; D012261:Ribonucleoproteins; D017411:Ribonucleoproteins, Small Nuclear; C028841:erythrocyte membrane band 4.1 protein; C120557:erythrocyte membrane protein band 4.1-like 1; C073288:SRSF2 protein, human; D000068103:Serine-Arginine Splicing Factors; D003850:Deoxyribonuclease I; D012259:Ribonuclease, Pancreatic