tailieunhanh - Báo cáo khoa hoc:" The Drosophila Anion Exchanger (DAE) lacks a detectable interaction with the spectrin cytoskeleton"

Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: The Drosophila Anion Exchanger (DAE) lacks a detectable interaction with the spectrin cytoskeleton | Dubreuil et al. Journal of Negative Results in BioMedicine 2010 9 5 http content 9 1 5 RESEARCH JOURNAL OF NEGATIVE RESULTS IN BIOMEDICINE Open Access The Drosophila Anion Exchanger DAE lacks a detectable interaction with the spectrin cytoskeleton Ronald R Dubreuil 1 Amlan Das2 Christine Base1 and G Harper Mazock1 Abstract Background Current models suggest that the spectrin cytoskeleton stabilizes interacting ion transport proteins at the plasma membrane. The human erythrocyte anion exchanger AE1 was the first membrane transport protein found to be associated with the spectrin cytoskeleton. Here we evaluated a conserved anion exchanger from Drosophila DAE as a marker for studies of the downstream effects of spectrin cytoskeleton mutations. Results Sequence comparisons established that DAE belongs to the SLC4A1-3 subfamily of anion exchangers that includes human AE1. Striking sequence conservation was observed in the C-terminal membrane transport domain and parts of the N-terminal cytoplasmic domain but not in the proposed ankyrin-binding site. Using an antibody raised against DAE and a recombinant transgene expressed in Drosophila S2 cells DAE was shown to be a 136 kd plasma membrane protein. A major site of expression was found in the stomach acid-secreting region of the larval midgut. DAE codistributed with an infolded subcompartment of the basal plasma membrane of interstitial cells. However spectrin did not codistribute with DAE at this site or in anterior midgut cells that abundantly expressed both spectrin and DAE. Ubiquitous knockdown of DAE with dsRNA eliminated antibody staining and was lethal indicating that DAE is an essential gene product in Drosophila. Conclusions Based on the lack of colocalization and the lack of sequence conservation at the ankyrin-binding site it appears that the well-characterized interaction between AE1 and the spectrin cytoskeleton in erythrocytes is not conserved in Drosophila. The results establish a pattern in .