The CyIIIa cytoskeletal actin gene of the sea urchin Strongylocentrotus purpuratus is activated in late cleavage and expressed exclusively in the aboral ectoderm territory of the embryo. Previous gene transfer studies defined a 2.3 kb cis-regulatory region that is necessary and sufficient for correct temporal and spatial expression of a CyIIIa.CAT fusion gene. In this paper, a negative regulatory element within this region was identified that is required for repression of the CyIIIa gene in skeletogenic mesenchyme cells. The repression mediated by this regulatory element takes place after initial territorial specification. A cDNA clone encoding a DNA-binding protein with twelve Zn fingers (SpZ12-1) was isolated by probing an expression library with this cis-element. Deletion analysis of the SpZ12-1 protein confirmed that a DNA-binding domain is located within the Zn finger region. SpZ12-1 is the only DNA-binding protein in embryo nuclear extract that interacts with the specific cis-target sites required for repression of CyIIIa.CAT in skeletogenic mesenchyme and is likely to be the trans factor that mediates this repression.

Anderson
R.
,
Britten
R. J.
,
Davidson
E. H.
(
1994
)
Repeated sequence target sites for maternal DNA-binding proteins in genes activated in early sea urchin development.
Dev. Biol
163
,
11
18
Barberis
A.
,
Superti-Furga
G.
,
Busslinger
M.
(
1987
)
Mutually exclusive interaction of the CCAAT-binding factor and a displacement protein with overlapping sequences of a histone gene promoter.
Cell
50
,
347
359
Benson
S.
,
Sucov
H. M.
,
Stephens
L.
,
Davidson
E. H.
,
Wilt
F. H.
(
1987
)
A lineage-specific gene encoding a major matrix protein of the sea urchin embryo spicule. I. Authentication of the cloned gene and its developmental expression.
Dev. Biol
120
,
499
506
Berg
J. M.
(
1990
)
Zinc finger domains: hypotheses and current knowledge.
Ann. Rev. Biophys. Biophys. Chem
19
,
405
421
Calzone
F. J.
,
Theze
N.
,
Thiebaud
P.
,
Hill
R. L.
,
Britten
R. J.
,
Davidson
E. H.
(
1988
)
Developmental appearance of factors that bind specifically to cis -regulatory sequences of a gene expressed in the sea urchin embryo.
Genes Dev
2
,
1074
1088
Cameron
R. A.
,
Davidson
E. H.
(
1991
)
Cell-type specification during sea urchin development.
Trends Genet
7
,
212
218
Char
B. R.
,
Bell
J. R.
,
Dovala
J.
,
Coffman
J. A.
,
Harrington
M. J.
,
Becerra
J.C.
,
Davidson
E. H.
,
Calzone
F. J.
,
Maxson
R.
(
1993
)
SpOct, a gene encoding the major octamer-binding protein in sea urchin embryos: expression profile, evolutionary relationships, and DNA binding of expressed protein.
Dev. Biol
158
,
350
363
Chowdhury
K.
,
Deutsch
U.
,
Gruss
P.
(
1987
)
A multigene family encoding several ‘finger’ structures is present and differentially active in mammalian genomes.
Cell
48
,
771
778
Coffman
J. A.
,
Davidson
E. H.
(
1992
)
Expression of spatially regulated genes in the sea urchin embryo.
Curr. Opin. Genet. Dev
2
,
260
268
Cox
K. H.
,
Angerer
L. M.
,
Lee
J. J.
,
Britten
R. J.
,
Davidson
E. H.
,
Angerer
R. C.
(
1986
)
Cell lineage-specific programs of expression of multiple actin genes during sea urchin embryogenesis.
J. Mol. Biol
188
,
159
172
Davidson
E. H.
(
1989
)
Lineage-specific gene expression and the regulative capacities of the sea urchin embryo: a proposed mechanism.
Development
105
,
421
445
Flytzanis
C. N.
,
Britten
R. J.
,
Davidson
E. H.
(
1987
)
Ontogenic activatin of a fusion gene introduced into the sea urchin egg.
Proc. Natl. Acad. Sci. USA
84
,
151
155
Franks
R. R.
,
Anderson
R.
,
Moore
J. G.
,
Hough-Evans
B. R.
,
Britten
R. J.
,
Davidson
E. H.
(
1990
)
Competitive titration in living sea urchin embryos of regulatory factors required for expression of the CyIIIa actin gene.
Development
110
,
31
40
Franks
R. R.
,
Hough-Evans
B. R.
,
Britten
R. J.
,
Davidson
E. H.
(
1988
)
Spatially deranged though temporally correct expression of a Strongylocentrotus purpuratus actin gene fusion in transgenic embryos of a different sea urchin family.
Genes Dev
2
,
1
12
Hickey
R. J.
,
Boshar
M. F.
,
Crain
W. R.
(
1987
)
Transcription of three actin genes and a repeated sequence in isolated nuclei of sea urchin embryos.
Dev. Biol
124
,
215
225
Höög
C.
,
Calzone
F. J.
,
Cutting
A. E.
,
Britten
R. J.
,
Davidson.
E. H.
(
1991
)
Gene regulatory factors of the sea urchin embryo II. Two dissimilar proteins, P3A1 and P3A2, bind to the same target sites that are required for early territorial gene expression.
Development
112
,
351
364
Hough-Evans
B. R.
,
Franks
R. R.
,
Cameron
R. A.
,
Britten
R. J.
,
Davidson
E. H.
(
1987
)
Correct cell type-specific expression of a fusion gene injected into sea urchin eggs.
Dev. Biol
121
,
576
579
Hough-Evans
B. R.
,
Britten
R. J.
,
Davidson
E. H.
(
1988
)
Mosaic incorporation and regulated expression of an exogenous gene in the sea urchin embryo.
Dev. Biol
129
,
198
208
Hough-Evans
B. R.
,
Franks
R. R.
,
Zeller
R. W.
,
Britten
R. J.
,
Davidson
E. H.
(
1990
)
Negative spatial regulation of the lineage-specific CyIIIa actin gene in the sea urchin embryo.
Development
110
,
41
50
Killian
C. E.
,
Wilt
F. H.
(
1989
)
The accumulation and translation of a spicule matrix protein mRNA during sea urchin embryo development.
Dev. Biol
133
,
148
156
Livant
D.
,
Cutting
A.
,
Britten
R. J.
,
Davidson
E. H.
(
1988
)
Titration of the activity of a fusion gene in intact sea urchin embryos.
Proc. Natl. Acad. Sci. USA
85
,
7607
7611
Livant
D.
,
Hough-Evans
B. R.
,
Moore
J. G.
,
Britten
R. J.
,
Davidson
E. H.
(
1991
)
Differential stability of expression of similarly specifiedendogenous and exogenous genes in the sea urchin embryo.
Development
113
,
385
398
Margolin
J. F.
,
Friedman
J. R.
,
Meyer
W.K.-H.
,
Vissing
H.
,
Thiesen
H.-J.
,
Rauscher
F. J.
III
(
1994
)
Kruppel -associated boxes are potent transcriptional repression domains.
Proc. Natl. Acad. Sci. USA
91
,
4509
4513
McMahon
A. P.
,
Flytzanis
C. N.
,
Hough-Evans
B. R.
,
Katula
K. S.
,
Britten
R. J.
,
Davidson
E. H.
(
1985
)
Introduction of cloned DNA into sea urchin egg cytoplasm: replication and persistence during embryogenesis.
Dev. Biol
108
,
420
430
Pavletich
N. P.
,
Pabo
C. O.
(
1991
).
Zinc finger-DNA recognition: crystal structure of a Zif268-DNA complex at 2.1 Å.
Science
252
,
809
817
Pavletich
N. P.
,
Pabo
C. O.
(
1993
)
Crystal structure of a five-finger GLI-DNA complex: new perspective on zinc fingers.
Science
261
,
1701
1707
Pellegrino
G. R.
,
Berg
J. M.
(
1991
)
Identification and characterization of ‘zinc finger’ domains by the polymerase chain reaction.
Proc. Natl. Acad. Sci. USA
88
,
671
675
Ransick
R.
,
Ernst
S. J.
,
Britten
R. J.
,
Davidson
E. H.
(
1993
)
Whole mount in situ hybridization shows Endo 16 to be a marker for the vegetal plate territory in sea urchin embryos.
Mech. Dev
42
,
117
124
Rosenberg
U. B.
,
Schröder
C.
,
Preiss
A.
,
Kienlin
A.
,
Côte
S.
,
Riede
L.
,
Jäckle
H.
(
1986
)
Structural homology of the product of the Drosophila Kruppel gene with Xenopus transcription factor IIIA.
Nature
319
,
336
339
Shott
R. J.
,
Lee
J. J.
,
Britten
R. J.
,
Davidson
E. H.
(
1984
)
Differential expression of the actin gene family of Strongylocentrotus purpuratus.
Dev. Biol
101
,
295
306
Studier
F. W.
,
Moffat
B. A.
(
1986
)
Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes.
J. Mol. Biol
189
,
113
130
Sucov
H. M.
,
Hough-Evans
B. R.
,
Franks
R. R.
,
Britten
R. J.
,
Davidson
E. H.
(
1988
)
A regulatory domain that directs lineage-specific expression of a skeletal matrix protein gene in the sea urchin embryo.
Genes Dev
2
,
1238
1250
Theze
N.
,
Calzone
F. J.
,
Thiebaud
P.
,
Hill
R. L.
,
Britten
R. J.
,
Davidson
E. H.
(
1990
)
Sequence of the CyIIIa actin gene regulatory domain bound specifically by sea urchin embryo nuclear proteins.
Mol. Reprod. Dev
25
,
110
122
Vinson
C. R.
,
LaMarco
K. L.
,
Johnson
P. F.
,
Landschultz
W. H.
,
McKnight
S. L.
(
1988
)
In situ detection of sequence-specific DNA binding activity specified by a recombinant bacteriophage.
Genes Dev
2
,
801
806
Witzgall
R.
,
O'Leary
E.
,
Laef
A.
,
Önaldi
D.
,
Bonventre
J. V.
(
1994
)
The Kruppl e-associated box-A (KRAB-A) domain of zinc finger proteins mediates transcriptional repression.
Proc. Natl. Acad. Sci. USA
91
,
4514
4518
Yuh
C.-H.
,
Ransick
A.
,
Martinez
P.
,
Britten
R. J.
,
Davidson
E.H.
(
1994
)
Complexity and organization of DNA-protein interactions in the 5¢ regulatory region of an endoderm-specific marker gene in the sea urchin embryo.
Mech. Dev
47
,
165
186
This content is only available via PDF.