Skip to main content
Advertisement

Main menu

  • Home
  • Articles
    • Accepted manuscripts
    • Issue in progress
    • Latest complete issue
    • Issue archive
    • Archive by article type
    • Special issues
    • Subject collections
    • Cell Scientists to Watch
    • First Person
    • Sign up for alerts
  • About us
    • About JCS
    • Editors and Board
    • Editor biographies
    • Travelling Fellowships
    • Grants and funding
    • Journal Meetings
    • Workshops
    • The Company of Biologists
    • Journal news
  • For authors
    • Submit a manuscript
    • Aims and scope
    • Presubmission enquiries
    • Fast-track manuscripts
    • Article types
    • Manuscript preparation
    • Cover suggestions
    • Editorial process
    • Promoting your paper
    • Open Access
    • JCS Prize
    • Manuscript transfer network
    • Biology Open transfer
  • Journal info
    • Journal policies
    • Rights and permissions
    • Media policies
    • Reviewer guide
    • Sign up for alerts
  • Contacts
    • Contact JCS
    • Subscriptions
    • Advertising
    • Feedback
    • For library administrators
  • COB
    • About The Company of Biologists
    • Development
    • Journal of Cell Science
    • Journal of Experimental Biology
    • Disease Models & Mechanisms
    • Biology Open

User menu

  • Log in
  • Log out

Search

  • Advanced search
Journal of Cell Science
  • COB
    • About The Company of Biologists
    • Development
    • Journal of Cell Science
    • Journal of Experimental Biology
    • Disease Models & Mechanisms
    • Biology Open

supporting biologistsinspiring biology

Journal of Cell Science

  • Log in
Advanced search

RSS   Twitter  Facebook   YouTube  

  • Home
  • Articles
    • Accepted manuscripts
    • Issue in progress
    • Latest complete issue
    • Issue archive
    • Archive by article type
    • Special issues
    • Subject collections
    • Cell Scientists to Watch
    • First Person
    • Sign up for alerts
  • About us
    • About JCS
    • Editors and Board
    • Editor biographies
    • Travelling Fellowships
    • Grants and funding
    • Journal Meetings
    • Workshops
    • The Company of Biologists
    • Journal news
  • For authors
    • Submit a manuscript
    • Aims and scope
    • Presubmission enquiries
    • Fast-track manuscripts
    • Article types
    • Manuscript preparation
    • Cover suggestions
    • Editorial process
    • Promoting your paper
    • Open Access
    • JCS Prize
    • Manuscript transfer network
    • Biology Open transfer
  • Journal info
    • Journal policies
    • Rights and permissions
    • Media policies
    • Reviewer guide
    • Sign up for alerts
  • Contacts
    • Contact JCS
    • Subscriptions
    • Advertising
    • Feedback
    • For library administrators
Journal Articles
Nuclear proteins of the bovine esophageal epithelium. II. The NuMA gene gives rise to multiple mRNAs and gene products reactive with monoclonal antibody W1
T.K. Tang, C.J. Tang, Y.L. Chen, C.W. Wu
Journal of Cell Science 1993 104: 249-260;
T.K. Tang
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
C.J. Tang
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Y.L. Chen
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
C.W. Wu
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • Article
  • Info & metrics
  • PDF
Loading

Summary

Treatment of small cells derived from the basal layer of bovine esophageal epithelium, with Triton X-100, urea and sonication resulted in a nuclear residue that was used as an immunogen for generation of monoclonal antibodies directed against nuclear components. One such antibody, designated W1, was found to label the nuclei of all cells examined. In interphase cells, the target antigen of antibody W1 was diffusely distributed in the nucleus. During metaphase, however, the W1 antigen formed prominent crescents at the poles of the mitotic spindle, diminished gradually in anaphase, and finally redistributed into the regenerating daughter nuclei. Western blotting with antibody W1 yielded a prominent polypeptide of M(r) approximately 230,000. The amino acid sequence, deduced from the nucleotide sequence of several overlapping cDNA clones that span the entire coding region, revealed that the W1 polypeptide was identical to the Nuclear Mitotic Apparatus (NuMA) protein, with a long alpha-helical central core flanked by two nonhelical domains. Interestingly, most cDNA sequences were identical to each other, except for six sequence blocks which were either inserted or deleted in individual cDNA clones. Analysis of the cDNA sequences of various clones, coupled with polymerase chain reaction amplification of cellular mRNA and genomic Southern blotting with region-specific probes, all indicated that multiple mRNA species were present in U-251 human glioma cells, derived from alternative splicing of the RNA transcript from a single NuMA/W1 gene. Besides the predominant form of the mRNA giving rise to the polypeptide of M(r) approximately 230,000, two other forms of mRNA, which arise as a result of alternative splicing and which use different translation termination codons, may yield lower molecular weight polypeptide products. Consistent with this notion, polypeptides of M(r) approximately 195,000 and approximately 194,000 have been observed in this and other studies on the NuMA/W1 protein. These data suggest that multiple isoforms of the NuMA polypeptides generated by alternative mRNA splicing may play some important functions which remain to be characterized.

  • © 1993 by Company of Biologists

REFERENCES

    1. Benz E. J. Jr..,
    2. Tang T. K.,
    3. Baklouti F.,
    4. Huang S.,
    5. Cho J. and
    6. Marchesi V. T.
    (1991). Tissue specific selection of alternative spliced exons of the protein 4. 1 gene generates multiple isoforms with altered spectrin-actin binding domains. Blood 78, 1–.
    OpenUrlFREE Full Text
    1. Cohen C. and
    2. Parry D. A. D.
    (1986). Alpha-helical coiled coils-a widespread motif in proteins. Trends Biochem. Sci 11, 245–248
    OpenUrlCrossRefWeb of Science
    1. Cohen J. B.,
    2. Broz S. D. and
    3. Levinson A. D.
    (1989). Expression of the H-ras proto-oncogene is controlled by alternative splicing. Cell 58, 461–472
    OpenUrlCrossRefPubMedWeb of Science
    1. Compton D. A.,
    2. Yen T. J. and
    3. Cleveland D. W.
    (1991). Identification of novel centromere/kinetochore-associated proteins using monoclonal antibodies generated against human mitotic chromosome scaffolds. J. Cell Biol 112, 1083–1097
    OpenUrlAbstract/FREE Full Text
    1. Compton D. A.,
    2. Szilak I. and
    3. Cleveland D. W.
    (1992). Primary structure of NuMA, an intracellular protein that defines a novel pathway for segregation of proteins at mitosis. J. Cell Biol 116, 1395–1408
    OpenUrlAbstract/FREE Full Text
    1. Costa F. F.,
    2. Agre P.,
    3. Watkins P. C.,
    4. Winkelmann J. C.,
    5. Tang T. K.,
    6. John K. M.,
    7. Lux S. E. and
    8. Forget B. G.
    (1990). Linkage of dominant hereditary spherocytosis to the gene for the erythrocyte membrane-skeleton protein ankyrin. New Engl. J. Med 323, 1046–1050
    OpenUrlCrossRefPubMedWeb of Science
    1. Cyr J. L.,
    2. Pfister K. K.,
    3. Bloom G. S.,
    4. Slaughter C. A. and
    5. Brady S. T.
    (1991). Molecular genetics of kinesin light chains: generation of isoforms by alternative splicing. Proc. Nat. Acad. Sci. USA 88, 10114–10118
    OpenUrlAbstract/FREE Full Text
    1. Himmler A.,
    2. Prechsel D.,
    3. Kirschner M. W. and
    4. Martin D. W.
    (1989). Tau consists of a set of proteins with repeated C-terminal microtubule-binding domains and variable N-terminal domains. Mol. Cell Biol 9, 1381–1388
    OpenUrlAbstract/FREE Full Text
    1. Irminger-Finger I.,
    2. Layman R. A. and
    3. Goldstein L. S. B.
    (1990). Analysis of the primary sequence and microtubule-binding region of the Drosophila 205K MAP. J. Cell Biol 111, 2563–2572
    OpenUrlAbstract/FREE Full Text
    1. Kallajoki M.,
    2. Weber K. and
    3. Osborn M.
    (1991). A 210 kD nuclearmatrix protein is a functional part of the mitotic spindle; a microinjection study using SPN monoclonal antibodies. EMBO J 10, 3351–3362
    OpenUrlPubMedWeb of Science
    1. Karlinsey J.,
    2. Stamatoyannopoulos G. and
    3. Enver T.
    (1989). Simultaneous purification of DNA and RNA from small numbers of eukaryotic cells. Anal. Biochem 180, 303–306
    OpenUrlCrossRefPubMedWeb of Science
    1. Koenig R. J.,
    2. Lazar M. A.,
    3. Hodin R. A.,
    4. Brent G. A.,
    5. Larsen P. R.,
    6. Chin W. W. and
    7. Moore D. D.
    (1985). Inhibition of thyroid hormone activation by non-hormone binding c-erbA protein generated by alternative mRNA splicing. Nature 337, 659–661
    OpenUrl
    1. Lewis S. A.,
    2. Wang D. and
    3. Cowan N. J.
    (1988). Microtubule-associated protein MAP2 shares a microtubule-binding motif with tau protein. Science 242, 936–939
    OpenUrlAbstract/FREE Full Text
    1. Lyderson B. and
    2. Pettijohn D.
    (1980). Human-specific nuclear protein that associates with the polar region of the mitotic apparatus: distribution in a human/hamster hybrid cell. Cell 22, 489–499
    OpenUrlCrossRefPubMedWeb of Science
    1. Noble M.,
    2. Lewis S. A. and
    3. Cowan N. J.
    (1989). The microtubule binding domain of microtubule-associated protein MAP1B contains a repeated sequence motif unrelated to that of MAP2 and tau. J. Cell Biol 109, 3367–3376
    OpenUrlAbstract/FREE Full Text
    1. Price C. M. and
    2. Pettijohn D. E.
    (1986). Distribution of the nuclear mitotic apparatus protein (NuMA) during mitosis and nuclear assembly. Exp. Cell Res 166, 295–311
    OpenUrlCrossRefPubMedWeb of Science
    1. Sellitto C. and
    2. Kuriyama R.
    (1988). Distribution of a matrix component of the midbody during the cell cycle in Chinese hamster ovary cells. J. Cell Biol 106, 431–439
    OpenUrlAbstract/FREE Full Text
    1. Smith D. B. and
    2. Johnson K. S.
    (1988). Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione-S-transferase. Gene 67, 37–40
    OpenUrl
    1. Tang T. K.,
    2. Leto T. L.,
    3. Correas I.,
    4. Alonso M. A.,
    5. Marchesi V. T. and
    6. Benz E. J. Jr..
    (1988). Selective expression of an erythroid-specific isoform of protein 4. 1. Proc. Nat. Acad. Sci. USA 85, 3713–3717
    OpenUrlAbstract/FREE Full Text
    1. Tang T. K.,
    2. Qin Z.,
    3. Leto T.,
    4. Marchesi V. T. and
    5. Benz E. J.
    (1990). Heterogeneity of mRNA and protein products arising from the protein 4. 1 gene in erythroid and nonerythroid tissues. J. Cell Biol 110, 617–624
    OpenUrlAbstract/FREE Full Text
    1. Tang T. K.,
    2. Hong T.-M.,
    3. Lin C.-Y.,
    4. Lai M.-L.,
    5. Liu C. H. L.,
    6. Lo H.-J.,
    7. Wang M.-E.,
    8. Chen L. B.,
    9. Chen W.-T.,
    10. Ip W.,
    11. Lin D. C.,
    12. Lin J. J.-C.,
    13. Lin S.,
    14. Sun T.-T.,
    15. Wang E.,
    16. Wang J. L.,
    17. Wu R.,
    18. Wu C.-W. and
    19. Chien S.
    (1993). Nuclear proteins of the bovine esophageal epithelium. I. Monoclonal antibody W2 specifically reacts with condensed nuclei of differentiated superficial cells. J. Cell Sci 104, 237–247
    OpenUrlAbstract/FREE Full Text
    1. Tousson A.,
    2. Zeng C.,
    3. Brinkley B. R. and
    4. Valdiria M. M.
    (1991). Centrophilin: a novel mitotic spindle protein involved in microtubule nucleation. J. Cell Biol 112, 427–440
    OpenUrlAbstract/FREE Full Text
    1. Wang E.,
    2. Cairncross J. G. and
    3. Liem R. K. H.
    (1984). Identification of glial filament protein and vimentin in the same intermediate filament system in human glioma cells. Proc. Nat. Acad. Sci. USA 81, 2102–2106
    OpenUrlAbstract/FREE Full Text
    1. Weber B. L.,
    2. Westin E. H. and
    3. Clarke M. F.
    (1990). Differentiation of mouse erythroleukemia cells enhanced by alternatively spliced c-myb mRNA. Science 249, 1291–1293
    OpenUrlAbstract/FREE Full Text
    1. Yang C. H.,
    2. Lambie E. J. and
    3. Snyder M.
    (1992). NuMA: an unusually long coiled-coil related protein in the mammalian nucleus. J. Cell Biol 116, 1303–1317
    OpenUrlAbstract/FREE Full Text
    1. Yang J. T.,
    2. Layman R. A. and
    3. Goldstein L. S. B.
    (1989). A three domain structure of the kinesin heavy chain revealed by DNA sequence and microtubule-binding analysis. Cell 56, 879–889
    OpenUrlCrossRefPubMedWeb of Science
    1. Yen J.,
    2. Wisdom R. M.,
    3. Tratner I. and
    4. Verma I. M.
    (1991). An alternative spliced form of FosB is a negative regulator of transcriptional activation and transformation by Fos proteins. Proc. Nat. Acad. Sci. USA 88, 5077–5081
    OpenUrlAbstract/FREE Full Text
Previous ArticleNext Article
Back to top
Previous ArticleNext Article

This Issue

 Download PDF

Email

Thank you for your interest in spreading the word on Journal of Cell Science.

NOTE: We only request your email address so that the person you are recommending the page to knows that you wanted them to see it, and that it is not junk mail. We do not capture any email address.

Enter multiple addresses on separate lines or separate them with commas.
Nuclear proteins of the bovine esophageal epithelium. II. The NuMA gene gives rise to multiple mRNAs and gene products reactive with monoclonal antibody W1
(Your Name) has sent you a message from Journal of Cell Science
(Your Name) thought you would like to see the Journal of Cell Science web site.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Journal Articles
Nuclear proteins of the bovine esophageal epithelium. II. The NuMA gene gives rise to multiple mRNAs and gene products reactive with monoclonal antibody W1
T.K. Tang, C.J. Tang, Y.L. Chen, C.W. Wu
Journal of Cell Science 1993 104: 249-260;
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
Citation Tools
Journal Articles
Nuclear proteins of the bovine esophageal epithelium. II. The NuMA gene gives rise to multiple mRNAs and gene products reactive with monoclonal antibody W1
T.K. Tang, C.J. Tang, Y.L. Chen, C.W. Wu
Journal of Cell Science 1993 104: 249-260;

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Alerts

Please log in to add an alert for this article.

Sign in to email alerts with your email address

Article navigation

  • Top
  • Article
  • Info & metrics
  • PDF

Related articles

Cited by...

More in this TOC section

  • Involvement of actin filaments and integrins in the binding step in collagen phagocytosis by human fibroblasts
  • University administration
  • The cytoplasmic domain of the interleukin-6 receptor gp80 mediates its basolateral sorting in polarized madin-darby canine kidney cells
Show more Journal Articles

Similar articles

Other journals from The Company of Biologists

Development

Journal of Experimental Biology

Disease Models & Mechanisms

Biology Open

Advertisement

Cell scientist to watch: Janet Iwasa

Read our interview with molecular animator Janet Iwasa, where she talks about her transition from the wet lab, explains how animation can facilitate research and discusses the challenges of the field.


New funding scheme supports sustainable events

As part of our Sustainable Conferencing Initiative, we are pleased to announce funding for organisers that seek to reduce the environmental footprint of their event. The next deadline to apply for a Scientific Meeting grant is 26 March 2021.


Mole – The Corona files

“Despite everything, it's just incredible that we get to do science.”

Mole continues to offer his wise words to researchers on how to manage during the COVID-19 pandemic.


JCS and COVID-19

For more information on measures Journal of Cell Science is taking to support the community during the COVID-19 pandemic, please see here.

If you have any questions or concerns, please do not hestiate to contact the Editorial Office.

Articles

  • Accepted manuscripts
  • Issue in progress
  • Latest complete issue
  • Issue archive
  • Archive by article type
  • Special issues
  • Subject collections
  • Interviews
  • Sign up for alerts

About us

  • About Journal of Cell Science
  • Editors and Board
  • Editor biographies
  • Travelling Fellowships
  • Grants and funding
  • Journal Meetings
  • Workshops
  • The Company of Biologists

For Authors

  • Submit a manuscript
  • Aims and scope
  • Presubmission enquiries
  • Fast-track manuscripts
  • Article types
  • Manuscript preparation
  • Cover suggestions
  • Editorial process
  • Promoting your paper
  • Open Access
  • JCS Prize
  • Manuscript transfer network
  • Biology Open transfer

Journal Info

  • Journal policies
  • Rights and permissions
  • Media policies
  • Reviewer guide
  • Sign up for alerts

Contacts

  • Contact JCS
  • Subscriptions
  • Advertising
  • Feedback

Twitter   YouTube   LinkedIn

© 2021   The Company of Biologists Ltd   Registered Charity 277992