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PMID: 9128242 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Structural protein 4.1 in the nucleus of human cells: dynamic rearrangements during cell division.

The Journal of cell biology ·Vol. 137 ·No. 2 ·1997-04-21 ·Pages 275-89

Krauss SW, Larabell CA, Lockett S, Gascard P, Penman S, Mohandas N, Chasis JA

Abstract

Structural protein 4.1, first identified as a crucial 80-kD protein in the mature red cell membrane skeleton, is now known to be a diverse family of protein isoforms generated by complex alternative mRNA splicing, variable usage of translation initiation sites, and posttranslational modification. Protein 4.1 epitopes are detected at multiple intracellular sites in nucleated mammalian cells. We report here investigations of protein 4.1 in the nucleus. Reconstructions of optical sections of human diploid fibroblast nuclei using antibodies specific for 80-kD red cell 4.1 and for 4.1 peptides showed 4.1 immunofluorescent signals were intranuclear and distributed throughout the volume of the nucleus. After sequential extractions of cells in situ, 4.1 epitopes were detected in nuclear matrix both by immunofluorescence light microscopy and resinless section immunoelectron microscopy. Western blot analysis of fibroblast nuclear matrix protein fractions, isolated under identical extraction conditions as those for microscopy, revealed several polypeptide bands reactive to multiple 4.1 antibodies against different domains. Epitope-tagged protein 4.1 was detected in fibroblast nuclei after transient transfections using a construct encoding red cell 80-kD 4.1 fused to an epitope tag. Endogenous protein 4.1 epitopes were detected throughout the cell cycle but underwent dynamic spatial rearrangements during cell division. Protein 4.1 was observed in nucleoplasm and centrosomes at interphase, in the mitotic spindle during mitosis, in perichromatin during telophase, as well as in the midbody during cytokinesis. These results suggest that multiple protein 4.1 isoforms may contribute significantly to nuclear architecture and ultimately to nuclear function.

MeSH Terms
3T3 Cells Amino Acid Sequence Animals Carcinoma, Squamous Cell Cell Division Cell Line Cell Nucleus/chemistry Cytoskeletal Proteins DNA Replication Diploidy Epitopes/analysis Erythrocyte Membrane/chemistry Female Fibroblasts/chemistry,cytology Humans Membrane Proteins/analysis Mice Molecular Sequence Data Neuropeptides Nuclear Matrix Nuclear Proteins/analysis Peptides Proliferating Cell Nuclear Antigen/analysis RNA Splicing Ribonucleoproteins Serine-Arginine Splicing Factors Spindle Apparatus/chemistry Tumor Cells, Cultured Uterine Cervical Neoplasms
Chemicals
Cytoskeletal Proteins Epitopes Membrane Proteins Neuropeptides Nuclear Proteins Peptides Proliferating Cell Nuclear Antigen Ribonucleoproteins SRSF2 protein, mouse erythrocyte membrane band 4.1 protein erythrocyte membrane protein band 4.1-like 1 SRSF2 protein, human Serine-Arginine Splicing Factors
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Krauss S W
Life Sciences Division, University of California, Lawrence Berkeley National Laboratory, 94720, USA.
Larabell C A
Lockett S
Gascard P
Penman S
Mohandas N
Chasis J A
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.137.2.275
Published
1997-04-21
Pages
275-89
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2139783
Subset
IM
Grants
NIDDK NIH HHS · DK 32094 · United States
External Links
PubMed source DOI: 10.1083/jcb.137.2.275 PMC Full Text View journal details
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