Presentation Type
Poster
Discipline Track
Other
Developmental Biology
Abstract Type
Research/Clinical
Abstract
Background: Hox genes encode transcriptional factors that regulate the expression of specific target genes along the anterior-posterior axis determining the segment identity during embryonic development. These master genes are expressed in the same order in which are located in the third chromosome of Drosophila melanogaster. As it was previously reported the interaction of neighboring Hox proteins as Scr-Antp and Antp-Ubx and one of the mechanisms for Hox protein regulation is protein-protein interaction, we determined the heterodimer formation of Ubx with AbdA and AbdB as well as the homodimer formation of these proteins in living cells which in turn could shed light about gene regulation mechanisms of their target genes and phenotypic suppression during Drosophila development.
Methods: We used Bimolecular Fluorescent Complementation (BiFC) to determine protein interactions between Ubx-AbdA and AbdB, AbdA-AbdB as well as the homodimer formation of Ubx, AbdA and AbdB. We constructed expression vectors carrying the coding sequences of Ubx, AbdA and AbdB fused to the N-terminal and C-terminal halves of Venus (VN and VC, respectively). Expression vectors were co-transfected with pCAGmCherry in HEK293 cells and red (Cherry) and green (BiFC) fluorescent cells were quantified using imageJ to determine BiFC percentage.
Results: The results show that Ubx interacts with AbdA and with AbdB giving 80% and 86% of interaction, respectively and AbdB also interacts with AbdA giving 90% of interaction. Surprisingly, interactions of Ubx-AbdA and AbdB-AbdA are affected by BiFC topology. Our results also demonstrated that Ubx and AbdB forms homodimers giving 82% and 96% of interaction, while AbdA does not.
Conclusions: We found heterodimer interactions of Ubx with AbdA and AbdB as well as AbdA with AbdB. Also, we showed homodimers formation of Ubx and AbdB while AbdA does not form homodimers. Altogether, these Hox interactions increase the evidence of neighboring interactions between Hox proteins and suggest a similar regulation mechanism that open the possibility to study its effect on the regulation of their target genes as well as phenotypic suppression during Drosophila development.
Academic/Professional Position
Graduate Student
Recommended Citation
Méndez, Rubén Montalvo; Jiménez-Mejía, Gustavo; and Reséndez-Pérez, Diana, "Ultrabithorax, Abdominal-A and Abdominal-B form heterodimers and homodimers in living cells by BiFC" (2024). Research Symposium. 68.
https://scholarworks.utrgv.edu/somrs/2024/posters/68
Ultrabithorax, Abdominal-A and Abdominal-B form heterodimers and homodimers in living cells by BiFC
Background: Hox genes encode transcriptional factors that regulate the expression of specific target genes along the anterior-posterior axis determining the segment identity during embryonic development. These master genes are expressed in the same order in which are located in the third chromosome of Drosophila melanogaster. As it was previously reported the interaction of neighboring Hox proteins as Scr-Antp and Antp-Ubx and one of the mechanisms for Hox protein regulation is protein-protein interaction, we determined the heterodimer formation of Ubx with AbdA and AbdB as well as the homodimer formation of these proteins in living cells which in turn could shed light about gene regulation mechanisms of their target genes and phenotypic suppression during Drosophila development.
Methods: We used Bimolecular Fluorescent Complementation (BiFC) to determine protein interactions between Ubx-AbdA and AbdB, AbdA-AbdB as well as the homodimer formation of Ubx, AbdA and AbdB. We constructed expression vectors carrying the coding sequences of Ubx, AbdA and AbdB fused to the N-terminal and C-terminal halves of Venus (VN and VC, respectively). Expression vectors were co-transfected with pCAGmCherry in HEK293 cells and red (Cherry) and green (BiFC) fluorescent cells were quantified using imageJ to determine BiFC percentage.
Results: The results show that Ubx interacts with AbdA and with AbdB giving 80% and 86% of interaction, respectively and AbdB also interacts with AbdA giving 90% of interaction. Surprisingly, interactions of Ubx-AbdA and AbdB-AbdA are affected by BiFC topology. Our results also demonstrated that Ubx and AbdB forms homodimers giving 82% and 96% of interaction, while AbdA does not.
Conclusions: We found heterodimer interactions of Ubx with AbdA and AbdB as well as AbdA with AbdB. Also, we showed homodimers formation of Ubx and AbdB while AbdA does not form homodimers. Altogether, these Hox interactions increase the evidence of neighboring interactions between Hox proteins and suggest a similar regulation mechanism that open the possibility to study its effect on the regulation of their target genes as well as phenotypic suppression during Drosophila development.