We report on the characterization of the first loss-of-function mutation in a Drosophila CREB gene, dCREB-A. In the epidermis, dCREB-A is required for patterning cuticular structures on both dorsal and ventral surfaces since dCREB-A mutant larvae have only lateral structures around the entire circumference of each segment. Based on results from epistasis tests with known dorsal/ventral patterning genes, we propose that dCREB-A encodes a transcription factor that functions near the end of both the DPP- and SPI-signaling cascades to translate the corresponding extracellular signals into changes in gene expression. The lateralizing phenotype of dCREB-A mutants reveals a much broader function for CREB proteins than previously thought.

Abel
T.
,
Bhatt
R.
,
Maniatis
T.
(
1992
)
A Drosophila CREB/ATF transcriptional activator binds to both fat body-and liver-specific regulatory elements.
Genes Dev
6
,
466
480
Alberga
A.
,
Boulay
J.-L.
,
Kempe
E.
,
Dennefeld
C.
,
Haenlin
M.
(
1991
)
The snail gene required for mesoderm formation in Drosophila isexpressed dynamically in derivatives of all three germ layers.
Development
111
,
983
992
Andrew
D. J.
,
Horner
M. A.
,
Petitt
M. G.
,
Smolik
S. M.
,
Scott
M. P.
(
1994
)
Setting limits on homeotic gene function: Restraint of Sex combs reduced activity by teashirt and other homeotic genes.
EMBO J
13
,
1132
1144
Barton
K.
,
Muthusamy
N.
,
Chanyangam
M.
,
Fischer
C.
,
Clendenin
C.
,
Leiden
J. M.
(
1996
)
Defective thymocyte proliferation and IL-2 production in transgenic mice expressing a dominant-negative form of CREB.
Nature
379
,
81
85
Bartsch
D.
,
Ghirardi
M.
,
Skehel
P. A.
,
Karl
K. A.
,
Herder
S. P.
,
Chen
M.
,
Bailey
C. H.
,
Kandel
E. R.
(
1995
)
Aplysia CREB2 represses long-term facilitation: Relief of repression converts transient facilitation into long-term functional and structural changes.
Cell
83
,
979
992
Basler
K.
,
Struhl
G.
(
1994
)
Compartment boundaries and the control of Drosophila limb pattern by hedgehog protein.
Nature
368
,
208
214
Blendy
J. A.
,
Kaestner
K. H.
,
Weinbauer
G. F.
,
Nieschlag
E.
,
Schutz
G.
(
1996
)
Severe impairment of spermatogenesis in mice lacking the CREM gene.
Nature
380
,
162
165
Blendy
J. A.
,
Kaestner
K. H.
,
Schmid
W.
,
Gass
P.
,
Schutz
G.
(
1996
)
Targeting of the CREB gene leads to up-regulation of a novel CREB mRNA isoform.
EMBO J
15
,
1098
1106
Bourtchuladze
R.
,
Frenguelli
B.
,
Blendy
J.
,
Cioffi
D.
,
Schutz
G.
,
Silva
A. J.
(
1994
)
Deficient long-term memory in mice with a targeted mutation of the cAMP-responsive element-binding protein.
Cell
79
,
59
68
Boutillier
A.-L.
,
Barthel
F.
,
Roberts
J. L.
,
Loeffler
J.-P.
(
1992
)
-adrenergic stimulation of cFOS via protein kinase A is mediated by cAMP regulatory element binding protein (CREB)-dependent and tissue-specific CREB-independent mechanisms in corticotrope cells.
Journal of Biological Chemistry
267
,
23520
23526
Brummel
T. J.
,
Twombly
V.
,
Marques
G.
,
Wrana
J. L.
,
Newfeld
S. J.
,
Attisano
L.
,
Massague
J.
,
O'Connor
M. B.
,
Gelbart
W. M.
(
1994
)
Characterization and relationship of Dpp receptors encoded by the saxophone and thick veins genes in Drosophila.
Cell
78
,
251
261
Cadigan
K. M.
,
Grossniklaus
U.
,
Gehring
W. J.
(
1994
)
Localized expression of sloppy paired maintains the polarity of Drosophila parasegments.
Genes Dev
8
,
899
913
Chen
R.-H.
,
Sarnecki
C.
,
Blenis
J.
(
1992
)
Nuclear localization and regulation of erk- and rsk- encoded protein kinases.
Mol. Cell. Biol
12
,
915
927
Chrivia
J. C.
,
Kwok
R. P. S.
,
Lamb
N.
,
Hagiwara
M.
,
Montminy
M. R.
,
Goodman
R. H.
(
1993
)
Phosphorylated CREB binds specifically to the nuclear protein CBP.
Nature
365
,
855
859
Comb
M.
,
Birnberg
N. C.
,
Seasholtz
A.
,
Herbert
E.
,
Goodman
H. M.
(
1986
)
A cyclic AMP-and phorbol ester-inducible DNA element.
Nature
323
,
353
356
Dash
P. K.
,
Hochner
B.
,
Kandel
E. R.
(
1990
)
Injection of the cAMP-responsive element into the nucleus of Aplysia sensory neurons blocks long-term facilitation.
Nature
345
,
718
721
de Groot
R. P.
,
Ballou
L. M.
,
Sassone-Corsi
P.
(
1994
)
Positive regulation of the cAMP-responsive activator CREM by the p70 S6 kinase: An alternative route to mitogen-induced gene expression.
Cell
79
,
81
91
Delegeane
A. M.
,
Ferland
L. H.
,
Mellon
P. L.
(
1987
)
Tissue-specific enhancer of the human glycoprotein hormone-subunit gene: Dependence on cyclic AMP-inducible elements.
Mol. Cell. Biol
7
,
3994
4002
Du
W.
,
Maniatis
T.
(
1992
)
An ATF/CREB binding site protein is required for virus induction of the human interferongene.
Proc. Natl. Acad. Sci. USA
89
,
2150
2154
Dwarki
V. J.
,
Montminy
M.
,
Verma
I. M.
(
1990
)
Both the basic region and the ‘leucine zipper’ domain of the cyclic AMP response element binding (CREB) protein are essential for transcriptional activation.
EMBO J
9
,
225
232
Erikson
E.
,
Maller
J. S.
(
1988
)
Substrate specificity of ribosomal protein S6 kinase II from Xenopus eggs.
Second Messengers and Phosphoproteins
12
,
135
143
Finkelstein
R.
,
Smouse
D.
,
Capaci
T.
,
Spradling
A.
,
Perrimon
N.
(
1990
)
The orthodenticle gene encodes a novel homeodomain protein involved in the development of the Drosophila nervous system and ocellar visual structures.
Genes Dev
4
,
1516
1527
Gaul
U.
,
Seifert
E.
,
Schuh
R.
,
Jäckle
H.
(
1987
)
Analysis of Kruppel protein distribution during early Drosophila development reveals posttranscriptional processing.
Cell
50
,
639
647
Giniger
E.
,
Jan
L. Y.
,
Jan
Y. N.
(
1993
)
Specifying the path of the intersegmental nerve of the Drosophila embryo; A role for Delta and Notch.
Development
117
,
431
440
Ginty
D. D.
,
Bonni
A.
,
Greenberg
M. E.
(
1994
)
Nerve growth factor activates a Ras-dependent protein kinase that stimulates c- fos transcription via phosphorylation of CREB.
Cell
77
,
713
725
Ginty
D. D.
,
Kornhauser
J. M.
,
Thompson
M. A.
,
Bading
H.
,
Mayo
K. E.
,
Takahashi
J. S.
,
Greenberg
M. E.
(
1993
)
Regulation of CREB phosphorylation in the suprachiasmatic nucleus by light and a circadian clock.
Science
260
,
238
241
Goda
Y.
(
1995
)
Photographic memory in flies.
Current Biology
5
,
852
853
Grieder
N. C.
,
Nellen
D.
,
Burke
R.
,
Basler
K.
,
Affolter
M.
(
1995
)
schnurri is required for Drosophila Dpp signaling and encodes a zinc finger protein similar to the mammalian transcription factor PRDII-BF1.
Cell
81
,
791
800
Hagiwara
M.
,
Alberts
A.
,
Brindle
P.
,
Meinkoth
J.
,
Feramisco
J.
,
Deng
T.
,
Karin
M.
,
Shenolikar
S.
,
Montminy
M.
(
1992
)
Transcriptional attenuation following cAMP induction requires PP-1-mediated dephosphorylation of CREB.
Cell
70
,
105
113
Hardy
S.
,
Shenk
T.
(
1988
)
Adenoviral control regions activated by E1A and the cAMP response element bind to the same factor.
Proc. Natl. Acad. Sci. USA
85
,
4171
4175
Heemskerk
J.
,
DiNardo
S.
(
1994
)
Drosophilahedgehog acts as a morphogen in cellullar patterning.
Cell
76
,
449
461
Hoodless
P. A.
,
Haerry
T.
,
Abdollah
S.
,
Stapleton
M.
,
O'Connor
M. B.
,
Attisano
L.
,
Wrana
J. L.
(
1996
)
MADR1, a MAD-related protein that functions in BMP2 signaling pathways.
Cell
85
,
489
500
Hummler
E.
,
Cole
T. J.
,
Blendy
J. A.
,
Ganss
R.
,
Aguzzi
A.
,
Schmid
W.
,
Beermann
F.
,
Schutz
G.
(
1994
)
Targeted mutation of the CREB gene: Compensation within the CREB/ATF family of transcription factors.
Proc. Natl. Acad. Sci. USA
91
,
5647
5651
Hunter
T.
(
1995
)
Protein kinases and phosphatases: The yin and yang of protein phosphorylation and signaling.
Cell
80
,
225
236
Irish
V. F.
,
Gelbart
W. M.
(
1987
)
The decapentaplegic gene is required for dorsal-ventral patterning of the Drosophila embryo.
Genes Dev
1
,
868
879
Irvine
K. D.
,
Botas
J.
,
Jha
S.
,
Mann
R. S.
,
Hogness
D. S.
(
1993
)
Negative autoregulation by Ultrabithorax controls the level and pattern of its expression.
Development
117
,
387
399
Isaac
D. D.
,
Andrew
D. J.
(
1996
)
Tubulogenesis in Drosophila: a requirement for the trachealess gene product.
Genes Dev
10
,
103
117
Kramer
U. M.
,
Koornneef
I. W.
,
de Laat
S.
,
van den Einjden-van Raaij
A. J. M.
(
1991
)
TGF-1 induces phosphorylation of the cyclic AMP responsive element binding protein in ML-CC164 cells.
EMBO J
10
,
1083
1089
Kwok
R. P.
,
Lundblad
J. R.
,
Chrivia
J. C.
,
Richards
J. C.
,
Bachinger
H. P.
,
Brennan
R. G.
,
Roberts
S. G.
,
Green
M. R.
,
Goodman
R. H.
(
1994
)
Nuclear protein CBP is a coactivator for the transcription factor CREB.
Nature
370
,
223
226
Lee
K. A. W.
,
Masson
N.
(
1993
)
Transcriptional regulation by CREB and its relatives.
Biochim. Biophys. Acta
1174
,
221
233
Letsou
A.
,
Arora
K.
,
Wrana
J. L.
,
Simin
K.
,
Twombly
V.
,
Jamal
J.
,
Staehling-Hampton
K.
,
Hoffman
F. M.
,
Gelbart
W. M.
,
Massague
J.
,
O'Connor
M. B.
(
1995
)
Drosophila Dpp signaling is mediated by the punt gene product: A dual ligand-binding type II receptor of the TGFreceptor family.
Cell
80
,
899
908
Lin
Y.-S.
,
Green
M. R.
(
1988
)
Interaction of a common cellular transcription factor, ATF, with regulatory elements in both E1A-and cyclic AMP-inducible promoters.
Proc. Natl. Acad. Sci. USA
85
,
3396
3400
Liu
F.
,
Hata
A.
,
Baker
J. C.
,
Doody
J.
,
Cárcamo
J.
,
Harland
R.
,
Massague
J.
(
1996
)
A human Mad protein acting as a BMP-regulated transcriptional activator.
Nature
381
,
620
623
Lundblat
J. R.
,
Kwok
R. P. S.
,
Laurance
M. E.
,
Harter
M. L.
,
Goodman
R. H.
(
1995
)
Adenoviral E1A-associated protein p300 as a functional homologue of the transcriptional co-activator CBP.
Nature
374
,
85
88
Mayer
U.
,
Nusslein-Volhard
C.
(
1988
)
A group of genes required for pattern formation in the ventral ectoderm of the Drosophila embryo.
Genes Dev
2
,
1496
1511
McCormick
A.
,
Brady
H.
,
Theill
L. E.
,
Karin
M.
(
1990
)
Regulation of the pituitary-specific homeobox gene GHF1 by cell-autonomous and environmental cues.
Nature
345
,
829
832
Meyer
T. E.
,
Habener
J. F.
(
1993
)
Cyclic adenosine 3,5 -monophosphate response element binding protien (CREB) and related transcription-activating deoxyribonucleic acid-binding proteins.
Endocrine Reviews
14
,
269
290
Montminy
M. R.
,
Sevarino
K. A.
,
Wagner
J. A.
,
Mandel
G.
,
Goodman
R. H.
(
1986
)
Identification of a cyclic-AMP-resonsive element within the rat somatostatin gene.
Proc. Nat. Acad. Sci. USA
83
,
6682
6686
Nellen
D.
,
Affolter
M.
,
Basler
K.
(
1994
)
Receptor serine/threonine kinases implicated in the control of Drosophila body pattern by decapentaplegic.
Cell
78
,
225
237
Newfeld
S. J.
,
Chartoff
E. H.
,
Graff
J. M.
,
Melton
D. A.
,
Gelbart
W. M.
(
1996
)
Mothers against dpp encodes a conserved cytoplasmic protein required in DPP/TGF-responsive cells.
Development
122
,
2099
2108
Padgett
R. W.
,
St. Johnston R. D.
,
Gelbart
W. M.
(
1987
)
A transcript from a Drosophila pattern gene predicts a protein homologous to the transforming growth factor-beta family.
Nature
325
,
81
84
Pankratz
M. J.
,
Hoch
M.
(
1995
)
Control of epithelial morphogenesis by cell signaling and integrin molecules in the Drosophila foregut.
Development
121
,
1885
1898
Panzer
S.
,
Weigel
D.
,
Beckendorf
S. K.
(
1992
)
Organogenesis in Drosophila melanogaster: Embryonic salivary gland determination is controlled by homeotic and dorsoventral patterning genes.
Development
114
,
49
57
Patel
N. H.
,
Martin-Blanco
E.
,
Coleman
K. G.
,
Poole
S. J.
,
Ellis
M. C.
,
Kornberg
T. B.
,
Goodman
C. S.
(
1989
)
Expression of engrailed proteins in arthropods, annelids, and chordates.
Cell
58
,
955
968
Patel
N. H.
,
Snow
P. M.
,
Goodman
C. S.
(
1987
)
Characterization and cloning of fasciclin III: A glycoprotein expressed on a subset of neurons and axon pathways in Drosophila.
Cell
56
,
1085
1092
Penton
A.
,
Chen
Y.
,
Staehling-Hampton
K.
,
Wrana
J. L.
,
Attisano
L.
,
Szidonya
J.
,
Cassil
J. A.
,
Massague
J.
,
Hoffman
F. M.
(
1994
)
Identification of two bone morphogenetic protein type I receptors in Drosophila and evidence that BRK25D is a decapentaplegic receptor.
Cell
78
,
239
250
Peunova
N.
,
Enikolopov
G.
(
1993
)
Amplification of calcium-induced gene transcription by nitric oxide in neuronal cells.
Nature
364
,
450
453
Quinn
P.
(
1993
)
Distinct activation domains within cAMP response element-binding protein (CREB) mediate basal and cAMP-stimulated transcription.
J. Biol. Chem
268
,
16999
17009
Raftery
L. A.
,
Twombly
V.
,
Wharton
K.
,
Gelbart
W. M.
(
1995
)
Genetic screens to identify elements of the decapentaplegic signaling pathway in Drosophila.
Genetics
139
,
241
254
Raz
E.
,
Shilo
B.-Z.
(
1993
)
Establishment of ventral cell fates in the Drosophila embryonic ectoderm requires DER, the EGF receptor homolog.
Genes Dev
7
,
1937
1948
Reuter
R.
,
Panganiban
G. E. F.
,
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
Rio
D. C.
,
Laski
F. A.
,
Rubin
G. M.
(
1986
)
Identification and immunochemical analysis of biologically active Drosophila P element transposase.
Cell
44
,
21
32
Rivera
V. M.
,
Miranti
C. K.
,
Misra
R. P.
,
Ginty
D. D.
,
Chen
R.-H.
,
Blenis
J.
,
Greenberg
M. E.
(
1993
)
A growth factor-induced kinase phosphorylates the serum response factor at a site that regulates its DNA-binding activity.
Mol. Cell. Biol
13
,
6260
6273
Roth
S.
,
Stein
D.
,
Nusslein-Volhard
C.
(
1989
)
A gradient of nuclear localization of the dorsal protein determines dorsoventral pattern in the Drosophila embryo.
Cell
59
,
1189
1202
Rutledge
B. J.
,
Zhang
K.
,
Bier
E.
,
Jan
Y. N.
,
Perrimon
N.
(
1992
)
The Drosophila spitz gene encodes a putative EGF-like growth factor involved in dorsal-ventral axis formation and neurogenesis.
Genes Dev
6
,
1503
1517
Sassone-Corsi
P.
,
Visvader
J.
,
Ferland
L.
,
Mellon
P. L.
,
Verma
I. M.
(
1988
)
Induction of proto-oncogene fos transcription through the adenylate cyclase pathway: characterization of a cAMP-responsive element.
Genes Dev
2
,
1529
1538
Schuske
K.
,
Hooper
J. E.
,
Scott
M. P.
(
1994
)
patched overexpression causes a loss of wingless expression in Drosophila embryos.
Dev. Biol
164
,
300
311
Sekelsky
J. J.
,
Newfeld
S. J.
,
Raftery
L. A.
,
Chartoff
E. H.
,
Gelbart
W. M.
(
1995
)
Genetic characterization and cloning of Mothers against dpp, a gene required for decapentaplegic function in Drosophila melanogaster.
Genetics
139
,
1347
1358
Sheng
M.
,
Thompson
M. A.
,
Greenberg
M. E.
(
1991
)
CREB: A Ca2+-regulated transcription factor phosphorylated by calmodulin-dependent kinases.
Science
252
,
1427
1430
Shilo
B.-Z.
,
Raz
E.
(
1991
)
Developmental control by the Drosophila EGF receptor homolog DER.
Trends Genet
7
,
388
392
Smolik
S. M.
,
Rose
R. E.
,
Goodman
R. H.
(
1992
)
A cyclic AMP-responsive element-binding transcriptional activator in Drosophila melanogaster, dCREB-A, is a member of the leucine zipper family.
Mol. Cell Biol
12
,
4123
4131
Stehle
J. H.
,
Foulkes
N. S.
,
Molina
C. A.
,
Simonneaux
V.
,
Pevet
P.
,
Sassone-Corsi
P.
(
1993
)
Adrenergic signals direct rhythmic expression of transcriptional repressor CREM in the pineal gland.
Nature
365
,
314
320
Struthers
R. S.
,
Vale
W. W.
,
Arias
C.
,
Sawchenko
P. E.
,
Montminy
M. R.
(
1991
)
Somatotroph hypoplasia and dwarfism in transgenic mice expressing a non-phosphorylatable CREB mutant.
Nature
350
,
622
624
Tepass
U.
,
Theres
C.
,
Knust
E.
(
1990
)
crumbs encodes an EGF-like protein expressed on apical membranes of Drosophila epithelial cells and required for organization of epithelia.
Cell
61
,
787
799
Tepass
U.
,
Theres
C.
,
Knust
E.
(
1990
)
Phenotypic and developmental analysis of mutations at the crumbs locus, a gene required for the development of epithelia in Drosophila melanogaster.
Cell
61
,
787
799
Usui
T.
,
Smolik
S. M.
,
Goodman
R. H.
(
1993
)
Isolation of Drosophila CREB-B: A novel CRE-binding protein.
DNA and Cell Biology
12
,
589
595
van den Heuvel
M.
,
Harryman-Samos
C.
,
Klingensmith
J.
,
Perrimon
N.
,
Nusse
R.
(
1993
)
Mutations in the segment polarity genes wingless and porcupine impair secretion of the wingless protein.
EMBO J
12
,
5293
5302
van den Heuvel
M.
,
Nusse
R.
,
Johnston
P.
,
Lawrence
P. A.
(
1989
)
Distribution of the wingless gene product in Drosophila embryos: a protein involved in cell-cell communication.
Cell
59
,
739
749
Wieschaus
E.
,
Perrimon
N.
,
Finkelstein
R.
(
1992
)
orthodenticle activity is required for the development of medial structures in the larval and adult epidermis of Drosophila.
Development
115
,
801
811
Wodarz
A.
,
Hinz
U.
,
Engelbert
M.
,
Knust
E.
(
1995
)
Expression of Crumbs confers apical character on plasma membrane domains of ectodermal epithelia of Drosophila.
Cell
82
,
67
76
Xing
J.
,
Ginty
D. D.
,
Greenberg
M. E.
(
1996
)
Coupling of the Ras/MAPK pathway to gene activation by RSK2, a growth factor-regulated CREB kinase.
Science
273
,
959
963
Yin
J. C. P.
,
Wallach
J. S.
,
Del Vecchio
M.
,
Wilder
E. L.
,
Zhou
H.
,
Quinn
W. G.
,
Tully
T.
(
1994
)
Induction of a dominant negative CREB transgene specifically blocks long-term memory in Drosophila.
Cell
79
,
49
58
Yin
J. C. P.
,
Del Vecchio
M.
,
Zhou
H.
,
Tully
T.
(
1995
)
CREB as a memory modulator: Induced expression of a dCREB2 activator isoform enhances long-term memory in Drosophila.
Cell
81
,
107
115
Yin
J. C. P.
,
Wallach
J. S.
,
Wilder
E. L.
,
Klingensmith
J.
,
Dang
D.
,
Perrimon
N.
,
Zhou
H.
,
Tully
T.
,
Quinn
W. G.
(
1995
)
A Drosophila CREB/CREM homolog encodes multiple isoforms, including a cyclic AMP-dependent protein kinase-responsive transcriptional activator and antagonist.
Mol. Cell. Biol
15
,
5123
5130
Zhang
Y.
,
Fresquez
C.
,
Holmgren
R.
(
1994
)
Ectopic expression of either the Drosophilagooseberry-distal or proximal gene causes alterations of cell fate in the epidermis and central nervous system.
Development
120
,
1151
1161
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