Functional synthetic Antennapedia genes and the dual roles of YPWM motif and linker size in transcriptional activation and repression
- PMID: 21712439
- PMCID: PMC3141989
- DOI: 10.1073/pnas.1108686108
Functional synthetic Antennapedia genes and the dual roles of YPWM motif and linker size in transcriptional activation and repression
Abstract
Segmental identity along the anteroposterior axis of bilateral animals is specified by Hox genes. These genes encode transcription factors, harboring the conserved homeodomain and, generally, a YPWM motif, which binds Hox cofactors and increases Hox transcriptional specificity in vivo. Here we derive synthetic Drosophila Antennapedia genes, consisting only of the YPWM motif and homeodomain, and investigate their functional role throughout development. Synthetic peptides and full-length Antennapedia proteins cause head-to-thorax transformations in the embryo, as well as antenna-to-tarsus and eye-to-wing transformations in the adult, thus converting the entire head to a mesothorax. This conversion is achieved by repression of genes required for head and antennal development and ectopic activation of genes promoting thoracic and tarsal fates, respectively. Synthetic Antennapedia peptides bind DNA specifically and interact with Extradenticle and Bric-à-brac interacting protein 2 cofactors in vitro and ex vivo. Substitution of the YPWM motif by alanines abolishes Antennapedia homeotic function, whereas substitution of YPWM by the WRPW repressor motif, which binds the transcriptional corepressor Groucho, allows all proteins to act as repressors only. Finally, naturally occurring variations in the size of the linker between the homeodomain and YPWM motif enhance Antennapedia repressive or activating efficiency, emphasizing the importance of linker size, rather than sequence, for specificity. Our results clearly show that synthetic Antennapedia genes are functional in vivo and therefore provide powerful tools for synthetic biology. Moreover, the YPWM motif is necessary--whereas the entire N terminus of the protein is dispensable--for Antennapedia homeotic function, indicating its dual role in transcriptional activation and repression by recruiting either coactivators or corepressors.
Conflict of interest statement
The authors declare no conflict of interest.
Figures
Similar articles
-
Transcription factor TFIIEβ interacts with two exposed positions in helix 2 of the Antennapedia homeodomain to control homeotic function in Drosophila.PLoS One. 2018 Oct 15;13(10):e0205905. doi: 10.1371/journal.pone.0205905. eCollection 2018. PLoS One. 2018. PMID: 30321227 Free PMC article.
-
The YPWM motif links Antennapedia to the basal transcriptional machinery.Development. 2008 May;135(9):1669-79. doi: 10.1242/dev.018028. Epub 2008 Mar 26. Development. 2008. PMID: 18367556
-
spen encodes an RNP motif protein that interacts with Hox pathways to repress the development of head-like sclerites in the Drosophila trunk.Development. 1999 Dec;126(23):5373-85. doi: 10.1242/dev.126.23.5373. Development. 1999. PMID: 10556062
-
Antennapedia: The complexity of a master developmental transcription factor.Genesis. 2024 Feb;62(1):e23561. doi: 10.1002/dvg.23561. Epub 2023 Oct 13. Genesis. 2024. PMID: 37830148 Review.
-
The bithorax complex of Drosophila an exceptional Hox cluster.Curr Top Dev Biol. 2009;88:1-33. doi: 10.1016/S0070-2153(09)88001-0. Curr Top Dev Biol. 2009. PMID: 19651300 Review.
Cited by
-
Radiation-sensitising effects of antennapedia proteins (ANTP)-SmacN7 on tumour cells.Int J Mol Sci. 2013 Dec 11;14(12):24087-96. doi: 10.3390/ijms141224087. Int J Mol Sci. 2013. PMID: 24336110 Free PMC article.
-
A combination of activation and repression by a colinear Hox code controls forelimb-restricted expression of Tbx5 and reveals Hox protein specificity.PLoS Genet. 2014 Mar 20;10(3):e1004245. doi: 10.1371/journal.pgen.1004245. eCollection 2014 Mar. PLoS Genet. 2014. PMID: 24651482 Free PMC article.
-
The early history of the eye-antennal disc of Drosophila melanogaster.Genetics. 2022 May 5;221(1):iyac041. doi: 10.1093/genetics/iyac041. Genetics. 2022. PMID: 35460415 Free PMC article.
-
Transcription factor TFIIEβ interacts with two exposed positions in helix 2 of the Antennapedia homeodomain to control homeotic function in Drosophila.PLoS One. 2018 Oct 15;13(10):e0205905. doi: 10.1371/journal.pone.0205905. eCollection 2018. PLoS One. 2018. PMID: 30321227 Free PMC article.
-
Regulation of Silk Genes by Hox and Homeodomain Proteins in the Terminal Differentiated Silk Gland of the Silkworm Bombyx mori.J Dev Biol. 2016 May 25;4(2):19. doi: 10.3390/jdb4020019. J Dev Biol. 2016. PMID: 29615585 Free PMC article. Review.
References
-
- Gehring WJ. Homeo boxes in the study of development. Science. 1987;236:1245–1252. - PubMed
-
- Lewis EB. A gene complex controlling segmentation in Drosophila. Nature. 1978;276:565–570. - PubMed
-
- McGinnis W, Krumlauf R. Homeobox genes and axial patterning. Cell. 1992;68:283–302. - PubMed
-
- McGinnis W, Levine MS, Hafen E, Kuroiwa A, Gehring WJ. A conserved DNA sequence in homoeotic genes of the Drosophila Antennapedia and bithorax complexes. Nature. 1984;308:428–433. - PubMed
-
- Gehring WJ, Kloter U, Suga H. Evolution of the Hox gene complex from an evolutionary ground state. Curr Top Dev Biol. 2009;88:35–61. - PubMed
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Molecular Biology Databases