Advertisement
JOURNAL ARTICLES
Functional domains in the Deformed protein
A. Zhu, M.A. Kuziora
Development 1996 122: 1577-1587;

Summary

A chimeric protein consisting of Deformed with a substituted Abdominal-B homeodomain (Dfd/Abd-B) is used to identify protein domains outside the homeodomain that are required for regulatory activity in vivo. A series of deletion proteins were generated based on regions showing amino acid composition similar to known regulatory domains. Each mutant protein can influence regulation of homeotic genes in a manner distinct from the intact protein. Activity was also tested using promoter elements from empty spiracles and Distal-less, two genes known to be directly regulated by Abdominal-B. Removal of the acidic region and the C-tail region convert the chimera from a strong activator to a repressor of the Distal-less element, but had comparatively little effect on the activation of the empty spiracles element. Constructs without a third domain, the N domain, fail to show any regulatory activity. The N domain is the only domain of the Dfd/Abd-B protein which exhibits significant activation activity when fused to a heterologous DNA binding domain. Our results suggest transcriptional activity of the N domain can be modulated by the acidic and C-tail domains.

Reference

    1. Ali N.,
    2. Bienz M.
    (1991) Functional dissection of DrosophilaAbdominal-B protein. Mech. Dev 35, 55–64
    OpenUrlCrossRefPubMed
    1. Appel B.,
    2. Sakonju S.
    (1993) Cell-type specific mechanisms of transcriptional repression by the homeotic gene products UBX and ABD-A in Drosophila embryos. EMBO J 12, 1099–1109
    OpenUrlPubMedWeb of Science
    1. Bachiller D.,
    2. Macías A.,
    3. Duboule D.,
    4. Morata G.
    (1994) Conservation of a functional hierarchy between mammalian and insect Hox/HOM genes. EMBO J 13, 1930–1941
    OpenUrlPubMedWeb of Science
    1. Bellen H. J.,
    2. O'Kane C. J.,
    3. Wilson O.,
    4. Grossniklaus U.,
    5. Pearson R. K.,
    6. Gehring W. J.
    (1989) P-element-mediated enhancer detection: a versatile method to study development in Drosophila. Genes Dev 3, 1288–1300
    OpenUrlAbstract/FREE Full Text
    1. Casanova J.,
    2. Sanchez-Herrero E.,
    3. Morata G.
    (1986) Identification and characterization of a parasegment specific regulatory element of the Abdominal-B gene of Drosophila. Cell 47, 627–636
    OpenUrlCrossRefPubMedWeb of Science
    1. Chadwick R.,
    2. McGinnis W.
    (1987) Temporal and spatial distribution of transcripts from the Deformed gene of Drosophila. EMBO J 6, 779–789
    OpenUrlPubMedWeb of Science
    1. Chan S.-K.,
    2. Mann R. S.
    (1993) The segment identity functions of Ultrabithorax are contained within its homeodomain and carboxy-terminal sequences. Genes Dev 7, 796–811
    OpenUrlAbstract/FREE Full Text
    1. Cohen S. M.,
    2. Brönner G.,
    3. Kuttner F.,
    4. Jurgens G.,
    5. Jäckle H.
    (1989) Distal-less encodes a homeodomain protein required for limb development in Drosophila. Nature 338, 432–434
    OpenUrlCrossRefPubMedWeb of Science
    1. Desplan C.,
    2. Theis J.,
    3. O'Farrell P. H.
    (1988) The sequence specificity of homeodomain-DNA interaction. Cell 54, 1081–1090
    OpenUrlCrossRefPubMedWeb of Science
    1. Eghtedarzadeh M. K.,
    2. Henikoff S.
    (1986) Use of oligonucleotides to generate large deletions. Nucl. Acids Res 14, 5115–.
    OpenUrlFREE Full Text
    1. Ekker S. C.,
    2. Young K. E.,
    3. von Kessler D. P.,
    4. Beachy P.
    (1991) Optimal DNA sequence recognition by the Ultrabithorax homeodomain of Drosophila. EMBO J 10, 1179–1186
    OpenUrlPubMedWeb of Science
    1. Evans T.,
    2. Felsenfeld G.
    (1991) Trans-activation of a globin promoter in nonerythroid cells. Mol. Cell. Biol 11, 843–853
    OpenUrlAbstract/FREE Full Text
    1. Gibson G.,
    2. Gehring W. J.
    (1988) Head and thoracic transformation caused by ectopic expression of Antennapedia during Drosophila development. Development 102, 657–675
    OpenUrlAbstract/FREE Full Text
    1. Gibson G.,
    2. Schier A.,
    3. LeMotte P.,
    4. Gehring W. J.
    (1990) The specificities of Sex combs reduced and Antennapedia are defined by distinct portion of each protein that includes the homeodomain. Cell 62, 1087–1103
    OpenUrlCrossRefPubMedWeb of Science
    1. Gill G.,
    2. Pascal E.,
    3. Tseng Z. H.,
    4. Tjian R.
    (1994) A glutamine-rich hydrophobic patch in transcription factor Sp1 contacts the dTAFII110 component of the Drosophila TFIID complex and mediates transcriptional activation. Proc. Natl. Acad. Sci. USA 91, 192–196
    OpenUrlAbstract/FREE Full Text
    1. Halder G.,
    2. Callaerts P.,
    3. Gehring W. J.
    (1995) Induction of ectopic eyes by targeted expression of the eyeless gene in Drosophila. Science 267, 1788–1792
    OpenUrlAbstract/FREE Full Text
    1. Hall C. V.,
    2. Jacob P. E.,
    3. Ringold G. M.,
    4. Lee F.
    (1983) Expression and regulation of Escherichia coli lacZ gene fusions in mammalian cells. J. Mol. App. Genet 2, 101–109
    OpenUrlPubMed
    1. Hall M. N.,
    2. Johnson A. D.
    (1987) Homeo domain of the yeast repressor alpha 2 is a sequence specific DNA-binding domain but is not sufficient for repression. Science 237, 1007–1012
    OpenUrlAbstract/FREE Full Text
    1. Han K.,
    2. Levine M. S.,
    3. Manley J. L.
    (1989) Synergistic activation and repression of transcription by Drosophila homeobox proteins. Cell 56, 573–583
    OpenUrlCrossRefPubMedWeb of Science
    1. Hayes P. H.,
    2. Sato T.,
    3. Denell R. E.
    (1984) Homeosis in Drosophila: The Ultrabithorax larval syndrome. Proc. Natl. Acad. Sci. USA 81, 545–549
    OpenUrlAbstract/FREE Full Text
    1. Hoey T.,
    2. Levine M.
    (1988) Divergent homeo box proteins recognize similar DNA sequences in Drosophila. Nature 332, 858–861
    OpenUrlCrossRefPubMed
    1. Jiang J.,
    2. Hoey T.,
    3. Levine M.
    (1991) Autoregulation of a segmentation gene in Drosophila: combinatorial interaction of the even-skipped homeo box protein with a distal enhancer element. Genes Dev 5, 265–277
    OpenUrlAbstract/FREE Full Text
    1. John A.,
    2. Smith S. T.,
    3. Jaynes J. B.
    (1995) Inserting the Ftz homeodomain into engrailed creates a dominant transcriptional repressor that specifically turns off Ftz target genes in vivo. Development 121, 1801–1813
    OpenUrlAbstract
    1. Johnson F. B.,
    2. Krasnow M. A.
    (1990) Stimulation of transcription by an Ultrabithorax protein in vitro. Genes Dev 4, 1044–1052
    OpenUrlAbstract/FREE Full Text
    1. Jones B.,
    2. McGinnis W.
    (1993) The regulation of empty spiracles by Abdominal-B mediates an abdominal segment identity function. Genes Dev 7, 229–240
    OpenUrlAbstract/FREE Full Text
    1. Kissinger C. R.,
    2. Liu B.,
    3. Martin-Blanco E.,
    4. Kornberg T. B.,
    5. Pabo C.O.
    (1990). Crystal structure of an engrailed homeodomain-DNA complex at 2.8 Å resolution: a framework for understanding homeodomain-DNA interactions. Cell 63, 579–590
    OpenUrlCrossRefPubMedWeb of Science
    1. Ko H. S.,
    2. Fast P.,
    3. McBride W.,
    4. Staudt L. M.
    (1988) A human protein specific for the immunoglobin octamer DNA motiff contains a functional homeobox domain. Cell 55, 135–144
    OpenUrlCrossRefPubMedWeb of Science
    1. Krasnow M. A.,
    2. Saffman E. E.,
    3. Kornfeld K.,
    4. Hogness D. S.
    (1989) Transcriptional activation and repression by Ultrabithorax proteins in cultured Drosophila cells. Cell 57, 1031–1043
    OpenUrlCrossRefPubMedWeb of Science
    1. Kuziora M. A.,
    2. McGinnis W.
    (1988) Autoregulation of a Drosophila homeotic selector gene. Cell 55, 477–485
    OpenUrlCrossRefPubMedWeb of Science
    1. Kuziora M. A.,
    2. McGinnis W.
    (1989) A homeodomain substitution changes the regulatory specificity of the Deformed protein in Drosophila embryos. Cell 59, 563–571
    OpenUrlCrossRefPubMedWeb of Science
    1. Kuziora M. A.,
    2. McGinnis W.
    (1990) Altering the regulatory targets of the Deformed protein in Drosophila embryos by substituting the Abdominal-B homeodomain. Mech. of Dev 33, 83–93
    OpenUrlCrossRefPubMed
    1. Kuziora M. A.
    (1993) Abdominal-B protein isoforms exhibit distinct cuticular transformations and regulatory activities when ectopically expressed in Drosphila embryos. Mech. Dev 42, 125–137
    OpenUrlCrossRefPubMedWeb of Science
    1. Lamka M. L.,
    2. Boulet A. M.,
    3. Sakonju S.
    (1992) Ectopic expression of UBX and ABD-B proteins in Drosophila embryogenesis: competition, not a functional hierarchy, explains phenotypic suppression. Development 116, 841–854
    OpenUrlAbstract/FREE Full Text
    1. Lehming N.,
    2. Thanos D.,
    3. Brickman J. M.,
    4. Ma J.,
    5. Maniatis T.,
    6. Ptashne M.
    (1994) An HMG-like protein that can switch a transcriptional activator to a repressor. Nature 371, 175–179
    OpenUrlCrossRefPubMedWeb of Science
    1. Lin L.,
    2. McGinnis W.
    (1992) Mapping functional specificity in the Dfd and Ubx homeo domain of Drosophila melanogaster. Genes Dev 6, 1071–1081
    OpenUrlAbstract/FREE Full Text
    1. Lohs-Schardin M.,
    2. Cremer C.,
    3. Nusslein-Volhard C.
    (1979) A fate map of the larval epidermis of Drosophila melanogaster: Localized cuticle defects following irradiation of the blastoderm with an ultraviolet laser microbeam. Dev. Biol 73, 239–255
    OpenUrlCrossRefPubMedWeb of Science
    1. Maheswaran S.,
    2. Park S.,
    3. Bernard A.,
    4. Morris J. F.,
    5. Rauscher F. J. I.,
    6. Hill D. E.,
    7. Harber D. A.
    (1993) Physical and functional interaction between WT1 and p53 proteins. Proc. Natl. Acad. Sci. USA 90, 5100–5104
    OpenUrlAbstract/FREE Full Text
    1. Malicki J.,
    2. Schughart K.,
    3. McGinnis W.
    (1990). Mouse Hox 2.2 specifies thoracic segmental identity in Drosophila embryos and larvae. Cell 63, 961–967
    OpenUrlCrossRefPubMedWeb of Science
    1. Mann R. S.,
    2. Hogness D. S.
    (1990) Functional dissection of Ultrabithorax proteins in D. melanogaster. Cell 60, 597–610
    OpenUrlCrossRefPubMedWeb of Science
    1. McGinnis W.,
    2. Garber R. L.,
    3. Wirz J.,
    4. Kuroiwa A.,
    5. Gehring W. J.
    (1984) A homologous protein-coding sequence in Drosophila homeotic genes and its conservation in other metazoans. Cell 37, 403–408
    OpenUrlCrossRefPubMedWeb of Science
    1. McGinnis N.,
    2. Kuziora M. A.,
    3. McGinnis W.
    (1990). Human Hox-4.2 and Drosophila Deformed encode similar regulatory specificities in Drosophila embryos and larvae. Cell 63, 969–976
    OpenUrlCrossRefPubMedWeb of Science
    1. McGinnis W.,
    2. Krumlauf R.
    (1992) Homeobox genes and axial patterning. Cell 68, 283–302
    OpenUrlCrossRefPubMedWeb of Science
    1. Merrill V. K. L.,
    2. Turner F. R.,
    3. Kaufman T. C.
    (1987) A genetic and developmental analysis of mutations in the Deformed locus in Drosophila melanogaster. Dev. Biol 122, 379–395
    OpenUrlCrossRefPubMed
    1. Mohler J.,
    2. Mahaffey J. W.,
    3. Deutsch E.,
    4. Vani K.
    (1995) Control of Drosophila head segment identity by the bZIP homeotic gene cnc. Development 121, 237–247
    OpenUrlAbstract
    1. Moscovici C.,
    2. Moscovici G. M.,
    3. Jimenez H.,
    4. Lai M. M. C.,
    5. Hayman M. J.,
    6. Vogt P. K.
    (1977) Continuous tissue culture cell lines derived from chemically induced tumors of Japanese quail. Cell 11, 95–103
    OpenUrlCrossRefPubMedWeb of Science
    1. Otting G.,
    2. Qian Y.,
    3. Muller M.,
    4. Affolter M.,
    5. Gehring W.,
    6. Wuthrich K.
    (1988) Secondary structure determination for the Antennapedia homeodomain by nuclear magnetic resonance and evidence for a helix-turn-helix motif. EMBO J 7, 4305–4309
    OpenUrlPubMedWeb of Science
    1. Patel N. H.,
    2. Martin-Blanco E.,
    3. Coleman K. G.,
    4. Poole S. J.,
    5. Ellis M. C.,
    6. Kornberg T. B.,
    7. Goodman C. S.
    (1989) Expression of engrailed proteins in arthropods, annelids, and chordates. Cell 58, 955–968
    OpenUrlCrossRefPubMedWeb of Science
    1. Powers M. A.,
    2. Forbes D. J.
    (1994) Cytosolic factors in nuclear transport: What's importin?. Cell 79, 931–934
    OpenUrlCrossRefPubMedWeb of Science
    1. Regulski M.,
    2. McGinnis N.,
    3. Chadwick R.,
    4. McGinnis W.
    (1987) Developmental and molecular analysis of Deformed: A homeotic gene controlling Drosophila head development. EMBO J 6, 767–777
    OpenUrlPubMedWeb of Science
    1. Samson M.,
    2. Jackson-Grusby L.,
    3. Brent R.
    (1989) Gene activation and DNA binding by DrosophilaUbx and abd-A proteins. Cell 57, 1045–1052
    OpenUrlCrossRefPubMedWeb of Science
    1. Scott M.P.,
    2. Weiner A.
    (1984) Structural relationships among genes that control development: Sequence homology between the Antennapedia Ultrabithorax, and fushi tarazu loci of Drosophila. Proc. Natl. Acad. Sci. USA 81, 4115–4119
    OpenUrlAbstract/FREE Full Text
    1. Smith D. B.,
    2. Johnson K. S.
    (1988) Single step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene 67, 31–40
    OpenUrlCrossRefPubMedWeb of Science
    1. Spradling A. C.,
    2. Rubin G. M.
    (1982) Transposition of cloned P-elements into Drosophila germ line chromosomes. Science 218, 341–347
    OpenUrlAbstract/FREE Full Text
    1. Subramaniam V.,
    2. Bomze H. M.,
    3. Lopez A. J.
    (1994) Functional differences between Ultrabithorax protein isoforms in Drosophila melanogaster: Evidence from elimination, substitution and ectopic expression of specific isoforms. Genetics 136, 979–991
    OpenUrlAbstract/FREE Full Text
    1. Tjian R.,
    2. Maniatis T.
    (1994) Transcriptional activation: A complex puzzle with few easy pieces. Cell 77, 5–8
    OpenUrlCrossRefPubMedWeb of Science
    1. Vachon G.,
    2. Cohen B.,
    3. Pfeifle C.,
    4. McGuffin M. E.,
    5. Botas J.,
    6. Cohen S.M.
    (1992) Homeotic genes of the Bithorax complex repress limb development in the abdomen of the Drosophila embryo through the target gene Distal-less. Cell 71, 437–450
    OpenUrlCrossRefPubMedWeb of Science
    1. Van der Meer J. M.
    (1977) Optical clean and permanent whole mount preparation for phase contrast microscopy of cuticular structures of insect larvae. Dros. Inf. Serv 52, 160–.
    OpenUrl
    1. Winslow G. M.,
    2. Hayashi S.,
    3. Krasnow M.,
    4. Hogness D. S.,
    5. Scott M. P.
    (1989) Transcriptional activation by the Antennapedia and fushi tarazu proteins in cultured Drosophila cells. Cell 57, 1017–1030
    OpenUrlCrossRefPubMedWeb of Science
    1. Yang H.,
    2. Evans T.
    (1992) Distinct roles for the two GATA-1 finger protein domains. Mol. Cell. Biol 12, 4562–4570
    OpenUrlAbstract/FREE Full Text
    1. Zeng C.,
    2. Pinsonneault J. G. G.,
    3. McGinnis N.,
    4. McGinnis W.
    (1994) Deformed protein binding sites and cofactor binding sites are required for the function of a small segment-specific regulatory element in Drosophila embryos. EMBO J 13, 2362–2377
    OpenUrlPubMedWeb of Science
    1. Zeng W.,
    2. Andrew D. J.,
    3. Mathies M. A.,
    4. Horner M. A.,
    5. Scott M. P.
    (1993) Ectopic expression and function of the Antp and Scr homeotic genes: The N terminus of the homeo domain is critical to functional specificity. Development 118, 339–352
    OpenUrlAbstract
    1. Zhao J. J.,
    2. Lazzarini R. A.,
    3. Pick L.
    (1993). The mouse Hox-1.3 gene is functionally equivalent to the Drosophila Sex combs reduced gene. Genes Dev 7, 343–354
    OpenUrlAbstract/FREE Full Text
Back to top

 Download PDF

Email
JOURNAL ARTICLES
Functional domains in the Deformed protein
A. Zhu, M.A. Kuziora
Development 1996 122: 1577-1587;
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
Citation Tools
Alerts

Article navigation

Other journals from The Company of Biologists

Advertisement

Kathryn Virginia Anderson (1952-2020)

Developmental geneticist Kathryn Anderson passed away at home on 30 November 2020. Tamara Caspary, a former postdoc and friend, remembers Kathryn and her remarkable contribution to developmental biology.

Zooming into 2021

In a new Editorial, Editor-in-Chief James Briscoe and Executive Editor Katherine Brown reflect on the triumphs and tribulations of the last 12 months, and look towards a hopefully calmer and more predictable year.

Read & Publish participation extends worldwide

Over 60 institutions in 12 countries are now participating in our Read & Publish initiative. Here, James Briscoe explains what this means for his institution, The Francis Crick Institute. Find out more and view our full list of participating institutions.

Upcoming special issues

Imaging Development, Stem Cells and Regeneration
Submission deadline: 30 March 2021
Publication: mid-2021

The Immune System in Development and Regeneration
Guest editors: Florent Ginhoux and Paul Martin
Submission deadline: 1 September 2021
Publication: Spring 2022

Both special issues welcome Review articles as well as Research articles, and will be widely promoted online and at key global conferences.

Development presents...

Our successful webinar series continues into 2021, with early-career researchers presenting their papers and a chance to virtually network with the developmental biology community afterwards. Sign up to join our next session:

10 February
Time: 13:00 (GMT)
Chaired by: preLights