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. 1990 Mar 1;110(3):617–624. doi: 10.1083/jcb.110.3.617

Heterogeneity of mRNA and protein products arising from the protein 4.1 gene in erythroid and nonerythroid tissues

PMCID: PMC2116033  PMID: 2307701

Abstract

Immunologically cross-reactive isoforms of the cytoskeletal element protein 4.1 have been identified in many tissues in which they exhibit heterogeneity of molecular weight, abundance, and intracellular localization. To examine the basis for isoform production in erythroid and nonerythroid tissues, we have compared the structure and expression of cDNAs isolated from human erythroid and nonerythroid sources. We have encountered cDNAs representing many distinct mRNA sequences. These exhibit complete nucleotide sequence homology along most of their lengths. Differences were confined to five sequence blocks designated Motifs I-V, which were present or absent in each mRNA moiety. Motif I was expressed only in erythroid cells; it encodes 21 amino acids in a well-characterized spectrin/actin binding domain. Motif II, located near the COOH terminus of the 80-kD "erythroid" protein 4.1 molecule is present in the vast majority of transcripts from both erythroid and nonerythroid cells. Motifs IV and V alter the 5' untranslated region: simultaneous insertion of Motif IV and deletion of Motif V in the untranslated region inserts a new initiator methionine and establishes a contiguous open reading frame encoding a novel 135-kD protein 4.1 molecule. By immunochemical analysis we have identified the longer isoform in cells. Our results are most consistent with tissue-specific alternative mRNA splicing of transcripts of the protein 4.1 gene to yield numerous isoforms. These isoforms exhibit tissue specificity and alter strategic portions of the molecule. Moreover, we describe a novel high molecular weight form of protein 4.1 that arises by splicing events which allow translation at an upstream site.

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Selected References

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  1. Aster J. C., Welsh M. J., Brewer G. J., Maisel H. Identification of spectrin and protein 4.1-like proteins in mammalian lens. Biochem Biophys Res Commun. 1984 Mar 15;119(2):726–734. doi: 10.1016/s0006-291x(84)80311-3. [DOI] [PubMed] [Google Scholar]
  2. Baines A. J., Bennett V. Synapsin I is a spectrin-binding protein immunologically related to erythrocyte protein 4.1. 1985 May 30-Jun 5Nature. 315(6018):410–413. doi: 10.1038/315410a0. [DOI] [PubMed] [Google Scholar]
  3. Bennett V. The membrane skeleton of human erythrocytes and its implications for more complex cells. Annu Rev Biochem. 1985;54:273–304. doi: 10.1146/annurev.bi.54.070185.001421. [DOI] [PubMed] [Google Scholar]
  4. Cohen C. M., Foley S. F., Korsgren C. A protein immunologically related to erythrocyte band 4.1 is found on stress fibres on non-erythroid cells. Nature. 1982 Oct 14;299(5884):648–650. doi: 10.1038/299648a0. [DOI] [PubMed] [Google Scholar]
  5. Conboy J. G., Chan J., Mohandas N., Kan Y. W. Multiple protein 4.1 isoforms produced by alternative splicing in human erythroid cells. Proc Natl Acad Sci U S A. 1988 Dec;85(23):9062–9065. doi: 10.1073/pnas.85.23.9062. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Conboy J., Kan Y. W., Shohet S. B., Mohandas N. Molecular cloning of protein 4.1, a major structural element of the human erythrocyte membrane skeleton. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9512–9516. doi: 10.1073/pnas.83.24.9512. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Conboy J., Mohandas N., Tchernia G., Kan Y. W. Molecular basis of hereditary elliptocytosis due to protein 4.1 deficiency. N Engl J Med. 1986 Sep 11;315(11):680–685. doi: 10.1056/NEJM198609113151105. [DOI] [PubMed] [Google Scholar]
  8. Correas I., Leto T. L., Speicher D. W., Marchesi V. T. Identification of the functional site of erythrocyte protein 4.1 involved in spectrin-actin associations. J Biol Chem. 1986 Mar 5;261(7):3310–3315. [PubMed] [Google Scholar]
  9. Correas I., Speicher D. W., Marchesi V. T. Structure of the spectrin-actin binding site of erythrocyte protein 4.1. J Biol Chem. 1986 Oct 5;261(28):13362–13366. [PubMed] [Google Scholar]
  10. Davies G. E., Cohen C. M. Platelets contain proteins immunologically related to red cell spectrin and protein 4.1. Blood. 1985 Jan;65(1):52–59. [PubMed] [Google Scholar]
  11. Goodman S. R., Casoria L. A., Coleman D. B., Zagon I. S. Identification and location of brain protein 4.1. Science. 1984 Jun 29;224(4656):1433–1436. doi: 10.1126/science.6374897. [DOI] [PubMed] [Google Scholar]
  12. Granger B. L., Lazarides E. Membrane skeletal protein 4.1 of avian erythrocytes is composed of multiple variants that exhibit tissue-specific expression. Cell. 1984 Jun;37(2):595–607. doi: 10.1016/0092-8674(84)90390-8. [DOI] [PubMed] [Google Scholar]
  13. Holt G. D., Haltiwanger R. S., Torres C. R., Hart G. W. Erythrocytes contain cytoplasmic glycoproteins. O-linked GlcNAc on Band 4.1. J Biol Chem. 1987 Nov 5;262(31):14847–14850. [PubMed] [Google Scholar]
  14. Kozak M. Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs. Nucleic Acids Res. 1984 Jan 25;12(2):857–872. doi: 10.1093/nar/12.2.857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Leto T. L., Pratt B. M., Madri J. A. Mechanisms of cytoskeletal regulation: modulation of aortic endothelial cell protein band 4.1 by the extracellular matrix. J Cell Physiol. 1986 Jun;127(3):423–431. doi: 10.1002/jcp.1041270311. [DOI] [PubMed] [Google Scholar]
  16. Marchesi V. T. Stabilizing infrastructure of cell membranes. Annu Rev Cell Biol. 1985;1:531–561. doi: 10.1146/annurev.cb.01.110185.002531. [DOI] [PubMed] [Google Scholar]
  17. Ngai J., Stack J. H., Moon R. T., Lazarides E. Regulated expression of multiple chicken erythroid membrane skeletal protein 4.1 variants is governed by differential RNA processing and translational control. Proc Natl Acad Sci U S A. 1987 Jul;84(13):4432–4436. doi: 10.1073/pnas.84.13.4432. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Spiegel J. E., Beardsley D. S., Southwick F. S., Lux S. E. An analogue of the erythroid membrane skeletal protein 4.1 in nonerythroid cells. J Cell Biol. 1984 Sep;99(3):886–893. doi: 10.1083/jcb.99.3.886. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Tang T. K., Leto T. L., Correas I., Alonso M. A., Marchesi V. T., Benz E. J., Jr Selective expression of an erythroid-specific isoform of protein 4.1. Proc Natl Acad Sci U S A. 1988 Jun;85(11):3713–3717. doi: 10.1073/pnas.85.11.3713. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Tchernia G., Mohandas N., Shohet S. B. Deficiency of skeletal membrane protein band 4.1 in homozygous hereditary elliptocytosis. Implications for erythrocyte membrane stability. J Clin Invest. 1981 Aug;68(2):454–460. doi: 10.1172/JCI110275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Zinn K., DiMaio D., Maniatis T. Identification of two distinct regulatory regions adjacent to the human beta-interferon gene. Cell. 1983 Oct;34(3):865–879. doi: 10.1016/0092-8674(83)90544-5. [DOI] [PubMed] [Google Scholar]

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