The products of the homeotic genes in Drosophila are transcription factors that are necessary to impose regional identity along the anterior-posterior axis of the developing embryo. However, the target genes under homeotic regulation that control this developmental process are largely unknown. We have utilized an immunopurification method to clone target genes of the Antennapedia protein (ANTP). We present here the characterization of centrosomin (cnn), one of the target genes isolated using this approach. The spatial and temporal expression of the cnn gene in the developing visceral mesoderm (VM) of the midgut and the central nervous system (CNS) of wild-type and homeotic mutant embryos is consistent with the idea that cnn is a homeotic target. In the VM, Antp and abdominal-A (abd-A) negatively regulate cnn, while Ultrabithorax (Ubx) shows positive regulation. In the CNS, cnn is regulated positively by Antp and negatively by Ubx and abd-A. Characterization of a cDNA encoding CNN predicts a novel structural protein with three leucine zipper motifs and several coiled-coil domains exhibiting limited homology to the rod portion of myosin. Immunocytochemical results demonstrate that the cnn encoded protein is localized to the centrosome and the accumulation pattern is coupled to the nuclear and centrosome duplication cycles of cleavage. In addition, evidence suggests that the expression of the cnn gene in the VM correlates with the morphogenetic function of Ubx in that tissue, i.e., the formation of the second midgut construction. The centrosomal localization of CNN and the involvement of microtubules in midgut morphogenesis suggest that this protein may participate in mitotic spindle assembly and the mechanics of morphogenesis through an interaction with microtubules, either directly or indirectly.

Abbott
M. K.
,
Kaufman
T. C.
(
1986
)
The relationship between the functional complexity and the molecular organization of the Antennapedia locus of Drosophila melanogaster.
Genetics
114
,
919
942
Andrew
D. J.
,
Scott
M. P.
(
1992
)
Downstream of the homeotic genes.
New Biol
4
,
5
15
Bieber
A.J.
,
Snow
P. M.
,
Hortsch
M.
,
Patel
N. H.
,
Jacobs
J. R.
,
Traquina
Z. R.
,
Schilling
J.
,
Goodman
C. S.
(
1989
)
Drosophila neuroglian: a member of the immunoglobulin superfaminly with extensive homology to the vertebrate neural adhesion molecule L1.
Cell
59
,
447
460
Blochlinger
K.
,
Bodmer
R.
,
Jan
L. Y.
,
Jan
Y. N.
(
1990
)
Patterns of expression of Cut, a protein required for external sensory organ development, in wild type and cut mutant Drosophila embryos.
GenesDev
4
,
1322
1331
Botas
J.
(
1993
)
Control of morphogenesis and differentiation by HOM/ Hox genes.
Curr. Opin. Cell Biol
5
,
1015
1022
Brand
A. H.
,
Perrimon
N.
(
1993
)
Targeted gene expression as a means of altering cell fates and generating dominant phenotypes.
Development
118
,
401
415
Campos-Ortega
J. A.
(
1990
)
Mechanisms of a cellular decision during embryonic development of Drosophila melanogaster: epidermogenesis or neurogenesis.
Adv. Genet
27
,
403
453
Capovilla
M.
,
Brandt
M.
,
Botas
J.
(
1994
)
Direct regulation of decapentaplegic by Ultrabithorax and its role in Drosophila midgut morphogenesis.
Cell
76
,
461
475
Cavener
D. R.
(
1987
)
Comparison of the consensus sequence flanking translational start sites in Drosophila and vertebrates.
Nucl. Acids Res
15
,
1353
1361
Chouinard
S.
,
Kaufman
T. C.
(
1991
)
Control of expression of thehomeotic labial (lab) locus of Drosophila melanogaster: evidence for both positive and negative autogenous regulation.
Development
113
,
1267
1280
Diederich
R. J.
,
Merrill
V. K. L.
,
Pultz
M. A.
,
Kaufman
T. C.
(
1989
)
Isolation, structure and expression of labial, a homeotic gene of the Antennapedia complex involved in Drosophila head development.
Genes Dev
3
,
399
414
Doe
C. Q.
,
Scott
M. P.
(
1988
)
Segmentation and homeotic gene function in the developing nervous system of Drosophila.
Trends Neurosci
11
,
101
106
Doxsey
S.J.
,
Stein
P.
,
Evans
L.
,
Calarco
P. D.
,
Kirschner
M.
(
1994
)
Pericentrin, a highly conserved centrosome protein involved in microtubule organization.
Cell
76
,
639
650
Gould
A. P.
,
White
R. A. H.
(
1992
)
Connectin, a target of homeotic gene control in Drosophila.
Development
116
,
1163
1174
Greig
S.
,
Akam
M.
(
1993
)
Homeotic genes autonomously specify one aspect of pattern in the Drosophila mesoderm.
Nature
362
,
630
632
Guo
L. H.
,
Stepien
P. P.
,
Brousseau
R.
,
Narang
S.
,
Thomas
D. Y.
,
Wu
R.
(
1984
)
Synthesis of human insulin gene. VIII. Construction of expression vectors for fused proinsulin production in E.coli.
Gene
12
,
251
254
Hoppier
S.
,
Bienz
M.
(
1994
)
Specification of a single cell type by a Drosophila homeotic gene.
Cell
76
,
689
702
Hortsch
M.
,
Bieber
A. J.
,
Patel
N. H.
,
Goodman
C. S.
(
1990
)
Differential splicing generates a nervous system specific form of Drosophila neuroglian.
Neuron
4
,
697
709
Hursh
D. A.
,
Padgett
R. W.
,
Gelbart
W. M.
(
1993
)
Cross-regulation of decapentaplegic and Ultrabithorax transcription in the embryonic visceral mesoderm of Drosophila.
Development
117
,
1211
1222
Immergluck
K.
,
Lawrence
P. A.
,
Bienz
M.
(
1990
)
Induction across germ layers in Drosophila mediated by a genetic cascade.
Cell
62
,
261
268
Karch
F.
,
Weiffenbach
B.
,
Peifer
M.
,
Bender
W.
,
Duncan
I.
,
Celniker
S.
,
Crosby
M.
,
Lewis
E. B.
(
1985
)
The abdominal region of the Bithorax complex.
Cell
43
,
81
96
Kaufman
T. C.
,
Seeger
M. A.
,
Olsen
G.
(
1990
)
Molecular and genetic organization of the Antennapedia gene complex of Drosophila melanogaster.
Adv. Genet
27
,
309
362
Kuriyama
R.
,
Nislow
C.
(
1992
)
Molecular components of the mitotic spindle.
BioEssays
14
,
81
88
Landschulz
W. H.
,
Johnson
P.G.
,
McKnight
S. L.
(
1988
)
The leucine zipper: A hypothetical structure common to a new class of DNA binding proteins.
Science
240
,
1759
1763
Lasko
P. F.
,
Pardue
M. L.
(
1988
)
Studies of the genetic organization of the vestigial microregion of Drosophila melanogaster.
Genetics
120
,
495
502
Levis
R.
,
Hazelrigg
T.
,
Rubin
G. M.
(
1985
)
Separable cis-acting control elements for expression of the white gene of Drosophila.
EMBO J
4
,
3489
3499
Lewis
E. B.
(
1978
)
A gene complex controlling segmentation in Drosophila.
Nature
276
,
265
270
Lindsley
D.L.
,
Zimm
G.
(
1987
)
The genome of Drosophila melanogaster. Part 3: Rearrangements.
Dros. Inf. Serv
65
,
1
203
Lupas
A.
,
VanDyke
M.
,
Stock
J.
(
1991
)
Predicting coiled-coils from protein sequences.
Science
252
,
1162
1164
Mahaffey
J. W.
,
Kaufman
T. C.
(
1987
)
Distribution of the Sex combs reduced gene products in Drosophila melanogaster.
Genetics
117
,
51
60
Matthews
K. A.
,
Miller
D. F. B.
,
Kaufman
T. C.
(
1990
)
Functional implications of the unusual spatial distribution of a minor-tubulin isotype in Drosophila: a common thread among chordotonal ligaments, developing muscle and testis cyst cells.
Dev. Biol
137
,
171
183
Matthews
K. A.
,
Rees
D.
,
Kaufman
T. C.
(
1993
)
A functionally specialized-tubulin is required for oocyte meiosis and cleavage mitoses in Drosophila.
Development
117
,
977
991
McCormack
K.
,
Tanouye
M.A.
,
Iverson
L. E.
,
Lin
J.-W.
,
Ramaswami
M.
,
McCormack
T.
,
Campanelli
J. T.
,
Mathew
M. K.
,
Rudy
B.
(
1991
)
A role for hydrophobic residues in the voltage dependent gating of Shaker K+channels.
Proc. Natl. Acad. Sci.USA
88
,
2931
2935
Mitchison
T.
,
Kirschner
M.
(
1984
)
Microtubule assembly nucleated by isolated centrosomes.
Nature
312
,
232
237
Nose
A.
,
Mahajan
V. B.
,
Goodman
C. S.
(
1992
)
Connectin: a homophilic cell adhesion molecule expressed on a subset of muscles and the motoneurons that innervate them in Drosophila.
Cell
70
,
553
567
O'Shea
E.
,
Klemm
J.
,
Kim
P.
,
Abler
T.
(
1991
)
X-ray structure of the GCN4 leucine zipper, a two stranded parallel, coiled coil.
Science
254
,
539
544
Panganiban
G. E.
,
Reuter
R.
,
Scott
M. P.
,
Hoffmann
F. M.
(
1990
)
A Drosophila growth factor homolog, decapentaplegic, regulates homeotic gene expression within and across germ layers during midgut morphogenesis.
Development
110
,
1041
1050
Ptashne
M.
(
1988
)
How eukaryotic transcriptional activators work.
Nature
335
,
683
689
Reuter
R.
,
Panganiban
G. E.
,
Hoffmann
F. M.
,
Scott
M. P.
(
1990
)
Homeotic genes regulate the spatial expression of putative growth factors in the visceral mesoderm of Drosophila embryos.
Development
110
,
1031
1040
Reuter
R.
,
Scott
M. P.
(
1990
)
Expression and function of the homoeotic genes Antennapedia and Sex combs reduced in the embryonic midgut of Drosophila.
Development
109
,
289
303
Sanchez-Herrero
E.
,
Vernos
I.
,
Marco
R.
,
Morata
G.
(
1985
)
Genetic organization of Drosophila bithorax complex.
Nature
313
,
108
113
Sanger
F.
,
Nicklen
S.
,
Coulson
A. R.
(
1977
)
DNA sequencing with chain terminating inhibitors.
Proc. Natl. Acad. Sci.USA
74
,
5463
5476
Sarge
K. D.
,
Zimarino
V.
,
Holm
K.
,
Wu
C.
,
Morimoto
R. I.
(
1991
)
Cloning and characterization of two mouse heat shock factors with distinct inducible and constitutive DNA binding ability.
Genes Dev
5
,
1902
1911
Simon
M. A.
,
Dodson
S. G.
,
Rubin
G. M.
(
1993
)
An SH3-SH2-SH3 protein is required for p21 Ras1 activation and binds to sevenless and Sos proteins in vitro.
Cell
73
,
169
177
Simonelig
M.
,
Anxolabehere
D.
(
1991
)
A P-element of Scaptomyza pallida is active in Drosophila melanogaster.
Proc. Natl. Acad. Sci.USA
88
,
6103
6107
Slack
J. M. W.
,
Holland
P. W. H.
,
Graham
C. F.
(
1993
)
The zootype and the phylotypic stage.
Nature
361
,
490
492
Tautz
D.
,
Pfeifle
C.
(
1989
)
A non-radioactive in situ hybridization method for the localization of specific RNAs in Drosophila embryos reveals translational control of the segmentation gene hunchback.
Chromosoma
98
,
81
85
Thorens
B.
,
Sarkar
H. K.
,
kaback
H. R.
,
Lodish
H.
(
1988
)
Cloning and functional expression in bacteria of a novel glucose transporter present in liver, intestine, kidney and-pancreatic islet cells.
Cell
55
,
281
290
Tremml
G.
,
Bienz
M.
(
1989
)
Homeotic gene expression in the visceral mesoderm of Drosophila embryos.
EMBO J
8
,
2677
2685
Vachon
G.
,
Cohen
B.
,
Pfeifle
C.
,
McGuffin
M. E.
,
Botas
J.
,
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
Wakimoto
B. T.
,
Kaufman
T. C.
(
1981
)
Analysis of larval segmentation in lethal genotypes associated with the Antennapedia gene complex in Drosophila melanogaster.
Dev. Biol
81
,
51
64
White
R. A.
,
Wilcox
M.
(
1984
)
Protein products of the bithorax complex in Drosophil a.
Cell
39
,
163
171
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