Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2004 Oct;168(2):877-93.
doi: 10.1534/genetics.104.029850.

CREB binding protein functions during successive stages of eye development in Drosophila

Justin P Kumar  1 , Tazeen Jamal, Alex Doetsch, F Rudolf Turner, Joseph B Duffy
Affiliations

CREB binding protein functions during successive stages of eye development in Drosophila

Justin P Kumar et al. Genetics. 2004 Oct.

Abstract

During the development of the compound eye of Drosophila several signaling pathways exert both positive and inhibitory influences upon an array of nuclear transcription factors to produce a near-perfect lattice of unit eyes or ommatidia. Individual cells within the eye are exposed to many extracellular signals, express multiple surface receptors, and make use of a large complement of cell-subtype-specific DNA-binding transcription factors. Despite this enormous complexity, each cell will make the correct developmental choice and adopt the appropriate cell fate. How this process is managed remains a poorly understood paradigm. Members of the CREB binding protein (CBP)/p300 family have been shown to influence development by (1) acting as bridging molecules between the basal transcriptional machinery and specific DNA-binding transcription factors, (2) physically interacting with terminal members of signaling cascades, (3) acting as transcriptional coactivators of downstream target genes, and (4) playing a key role in chromatin remodeling. In a screen for new genes involved in eye development we have identified the Drosophila homolog of CBP as a key player in both eye specification and cell fate determination. We have used a variety of approaches to define the role of CBP in eye development on a cell-by-cell basis.

PubMed Disclaimer

Figures

F<sc>igure</sc> 1.—
Figure 1.—
CBP interacts genetically with soD. Scanning electron micrographs of adult eyes are shown in A–E. Confocal images of third instar imaginal discs are shown in F–L. All genotypes are at top of each column. Red is F-actin; green is identified in each panel. Anterior is to the right. G4, GAL4.
F<sc>igure</sc> 2.—
Figure 2.—
Schematic of CBP variants. Each CBP variant is expressed within subdomains of the developing eye using the UAS/GAL4 misexpression system. See materials and methods for cloning strategies. Individual protein domains are coded.
F<sc>igure</sc> 3.—
Figure 3.—
CBP is expressed in all cells of the developing eye imaginal disc. Confocal images of third instar imaginal discs are shown. Genotypes are listed at the left of each row. (A and B) Low and high magnification view of CBP expression ahead of and behind the morphogenetic furrow (MF). (C) Schematic of cells within the eye disc. Gray circles represent cells ahead of the furrow. Red circles represent ommatidial clusters. Brown circles represent intervening cells. (D–F) CBP is present in cone (c) and photoreceptor cells (1–8). (G–I) CBP is present in the intervening cells. (G) CBP; (H) lz-GAL4/UAS-GFP; and (I) merge of G and H. Yellow arrows indicate intervening cells. Anterior is to the right.
F<sc>igure</sc> 4.—
Figure 4.—
CBP regulates eyes absent but not dachshund expression during eye development. Confocal images of third instar imaginal discs are shown. All genotypes are to the left of each row. (A–C) CBP and Eya are coexpressed. (D–F) Eya protein levels are reduced in CBP loss-of-function retinal clones. (G–I) CBP and Dac are coexpressed. (J–L) Dac protein levels are not regulated by CBP. Molecules visualized are listed in each panel. Arrows mark CBP loss-of-function clones. Arrowheads mark the morphogenetic furrow. Anterior is to the right.
F<sc>igure</sc> 5.—
Figure 5.—
Mutations within CBP alter dac, eya, and so expression in the embryonic visual system. Light microscope images of wild type (A, C, E, G, I, and K) and nejTC41 mutant embryos (B, D, F, H, J, and L) are shown. Genotypes are at the top of each column. Molecules visualized are listed to the left of each panel. Lateral views of embryos are shown in A, B, E, F, I, and J and dorsal views are shown in C, D, G, H, K, and L. Eyes Absent and Dachshund proteins are detected with antibodies. β-Gal antibodies were used to detect the pattern of so-lacZ. vp, visual primordium; pc, protocerebrum; m, mesoderm. Anterior is to the left.
F<sc>igure</sc> 6.—
Figure 6.—
CBP mutants inhibit eye development. (A and C) Scanning electron micrographs of adult eyes. (B and D) Light microscope sections of adult retinas. (E–J) Confocal images of third instar imaginal discs. Genotypes are listed to the left of each row. Wild-type eyes are shown in A and B. CBP clones are shown in C–J. Molecules visualized are listed within each panel. Red asterisk in D marks center of large clone that is devoid of photoreceptors. Red arrows in D mark ommatidia at the borders of clones. Yellow arrows in E–J mark position of CBP retinal mosaic clone. Note the reduction of Elav staining within the clone. Also note that Atonal expression within the clone remains normal. Anterior is to the right.
F<sc>igure</sc> 7.—
Figure 7.—
CBP mutants inhibit eye development. (A, C, and E) Scanning electron micrographs of adult eyes. (B and D) Light microscope sections of adult retinas. (F) Confocal image of third instar imaginal disc. Genotypes are listed to the left of each row. nej heteroallelic (nej131/nejP) eyes are shown in A and B. GMR-GAL4/UAS-dCBP RNAi is shown in C and D. ey-GAL4/UAS-dCBP RNAi is shown in E and F. Asterisk in D marks region of eye that is devoid of photoreceptors. Arrows in D mark ommatidia with malformed rhabdomeres. Anterior is to the right.
F<sc>igure</sc> 8.—
Figure 8.—
Expression of individual CBP variants results in specific effects on photoreceptor and cone cell development. (A, E, I, M, and Q) Confocal images of third instar eye discs. (B, F, J, and N) Confocal images of pupal retinas. (C, G, K, O, and R) Scanning electron micrographs of adult eyes. (D, H, L, P, and S) Light microscope sections of adult retinas. Genotypes are listed to the left of each row. In all cases the CBP variant is expressed from a UAS construct driven by GMR-GAL4. Eye discs are stained with an antibody against Elav. Pupal retinas are stained for F-actin. Yellow numbers mark the number of cone cells within an ommatidium. Yellow asterisk in S marks area that is devoid of photoreceptors. Yellow arrowheads mark photoreceptor clusters. Anterior is to the right.
F<sc>igure</sc> 9.—
Figure 9.—
CBP functions during R3, R4, and R7 photoreceptor cell specification. (A, C, E, G, and I) Scanning electron micrographs of adult eyes. (B, D, F, H, and J) Light microscope sections of adult retinas. Genotypes are listed at the sides of each row; G4 stands for GAL4. In all cases the CBP variant is expressed from a UAS construct driven by sev-GAL4. (A–H) White arrows mark ommatidia with two R3 cells. Diagonal stripe arrows mark ommatidia that have opposite chirality. Dotted arrows mark ommatidia in which the R3 cell has transformed into an R7. Horizontal stripe arrows mark ommatidia in which R4 has transformed into R7. Checkered arrows mark ommatidia in which R4 has not been specified. Orange arrow marks an ommatidium in which R7 has been deleted. Crosshatched arrows mark ommatidia in which both R3 and R4 have been transformed into R7, resulting in three R7 cells per cluster. (J) Plaid arrows mark the large outer photoreceptor that occupies the R7 cell position. Arrow key is at bottom right of figure.
F<sc>igure</sc> 10.—
Figure 10.—
Expression of CBP variant proteins ahead of the furrow inhibits eye development. Scanning electron micrographs of adult eyes are in shown A–C. Confocal images of third instar imaginal discs are shown in D–F. All genotypes are at the top of each column. In all cases the CBP variant is expressed from a UAS construct driven by ey-GAL4. Anterior is to the right.
F<sc>igure</sc> 11.—
Figure 11.—
Schematic of steps in eye development regulated by CBP. CBP has been shown to interact with the eye specification gene sine oculis and regulates the expression of eyes absent. These interactions happen ahead of the advancing morphogenetic furrow (purple text). Behind the morphogenetic furrow CBP functions during many stages of ommatidial assembly. Interestingly, the development of the R8 founder cell is not dependent upon CBP. We have demonstrated a strong requirement for CBP in the R3, R4, and R7 photoreceptors. We have yet to determine a requirement for CBP in the R2/R5 and R1/R6 pairs.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Ait-Si-Ali, S., D. Carlisi, S. Ramirez, L. C. Upegui-Gonzalez, A. Duquet et al., 1999. Phosphorylation by p44 MAP kinase/ERK1 stimulates CBP histone acetyl transferase activity in vitro. Biochem. Biophys. Res. Commun. 262: 157–162. - PubMed
    1. Akimaru, H., Y. Chen, P. Dai, D. X. Hou, M. Nonaka et al., 1997. a Drosophila CBP is a co-activator of cubitus interruptus in hedgehog signalling. Nature 386: 735–738. - PubMed
    1. Akimaru, H., D. X. Hou and S. Ishii, 1997. b Drosophila CBP is required for dorsal-dependent twist gene expression. Nat. Genet. 17: 211–214. - PubMed
    1. Arany, Z., W. R. Sellers, D. M. Livingston and R. Eckner, 1994. E1A-associated p300 and CREB-associated CBP belong to a conserved family of coactivators. Cell 77: 799–800. - PubMed
    1. Avantaggiati, M. L., V. Ogryzko, K. Gardner, A. Giordano, A. S. Levine et al., 1997. Recruitment of p300/CBP in p53-dependent signal pathways. Cell 89: 1175–1184. - PubMed

Publication types

MeSH terms