Abstract
In Drosophila melanogaster two high molecular weight tropomyosin isoforms, historically named heavy troponins (TnH-33 and TnH-34), are encoded by the Tm1 tropomyosin gene. They are specifically expressed in the indirect flight muscles (IFM). Their N-termini are conventional and complete tropomyosin sequences, but their C-termini consist of different IFM-specific domains that are rich in proline, alanine, glycine and glutamate. The evidence indicates that in Diptera these IFM-specific isoforms are conserved and are not troponins, but heavy tropomyosins (TmH). We report here that they are post-translationally modified by several phosphorylations in their C-termini in mature flies, but not in recently emerged flies that are incapable of flight. From stoichiometric measurements of thin filament proteins and interactions of the TmH isoforms with the standard Drosophila IFM tropomyosin isoform (protein 129), we propose that the TmH N-termini are integrated into the thin filament structural unit as tropomyosin dimers. The phosphorylated C-termini remain unlocated and may be important in IFM stretch-activation. Comparison of the Tm1 and Tm2 gene sequences shows a complete conservation of gene organisation in other Drosophilidae, such as Drosophila pseudoobscura, while in Anopheles gambiae only one exon encodes a single C-terminal domain, though overall gene organization is maintained. Interestingly, in Apis mellifera (hymenopteran), while most of the Tm1 and Tm2 gene features are conserved, the gene lacks any C-terminal exons. Instead these sequences are found at the 3’ end of the troponin I gene. In this insect order, as in Lethocerus (hemipteran), the original designation of troponin H (TnH) should be retained. We discuss whether the insertion of the IFM-specific pro-ala-gly-glu-rich domain into the tropomyosin or troponin I genes in different insect orders may be related to proposals that the IFM stretch activation mechanism has evolved independently several times in higher insects.
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Acknowledgements
We thank Dr Belinda Bullard for supplying the monoclonal antibodies, Dr. Jesús Vázquez and Anabel Marina for help with the MALDI-TOF analysis and Aida Villa for technical help. This research was supported by grants from the Spanish Government PB96-0069, ESP1999-0379-C02, ESP-2001-4521-PE, ESP2002-11913-E and ESP2003-09475-C02-01. JCS was funded by the BBSRC (UK). R. Herranz was supported during this work by a FPI fellowship of the Spanish Ministery of Science and Technology.
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MATEOS, J., HERRANZ, R., DOMINGO, A. et al. The structural role of high molecular weight tropomyosins in dipteran indirect flight muscle and the effect of phosphorylation. J Muscle Res Cell Motil 27, 189–201 (2006). https://doi.org/10.1007/s10974-005-9044-3
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DOI: https://doi.org/10.1007/s10974-005-9044-3