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Stretchin-klp, a novel Drosophila indirect flight muscle protein, has both myosin dependent and independent isoforms

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Abstract

Stretchin-klp is a newly described protein in Drosophila indirect flight muscles (IFM) that migrates on SDS gels as two distinct components of approximately 225 and 231 kD. Although the larger isoform is IFM specific, the smaller stretchin-klp isoform is expressed not only in IFM, but also in wild-type tissues of the adult head, abdomen and thorax from which the IFM has been removed. It is not detected, however, in jump or leg muscles. Probes derived from a cDNA encoding part of stretchin-klp hybridize with a 6.7 kb mRNA. Stretchin-klp is one of several putative products of the Stretchin-Myosin light chain kinase gene and is predicted to have multiple immunoglobulin domains arranged in tandem pairs separated by variable length spacers. Polyclonal antibodies directed against the expressed peptide of the stretchin-klp cDNA label the IFM myofibril A-band, though not its central and lateral regions. Analyses of IFM mutants indicate that the larger stretchin-klp isoform is myosin dependent. Although the normal adult myosin filament or the ‘headless’ myosin rod is sufficient for accumulation of both the large and small stretchin-klp isoforms, loss of myosin, or substitution of the adult rod with an embryonic one in IFM prevents the larger isoform from being formed or stabilized. During development stretchin-klp is first detected at pupal stage p8, when myofibrils are being constructed. These studies suggest that this newly identified protein is a major component of the Drosophila IFM thick filament.

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Acknowledgements

We are grateful to Bruce Hamilton for providing the λEXLOX adult Drosophila cDNA expression library. We thank Donald Gantz for advice with electron microscopy, Hector Lucero for guidance with molecular biology protocols, Marc Champagne for helpful discussions, Jeff Moore for comments on the manuscript and Sanford Bernstein, John Sparrow and Jim Vigoreaux for providing mutant stocks of Drosophila. Maria Ericsson (Harvard Medical School Core Electron Microscopy Facility) sectioned frozen preparations and provided technical assistance with immuno-gold staining. We are grateful to the Department of Physiology and Biophysics for supporting this work and to the former chairman of the Department of Physiology, the late Benjamin Kaminer, who was a generous, kind, encouraging mentor for over two decades.

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  1. Sunita R. Patel

    Present address: Department of Medicine, Division of Hematology, Brigham and Women’s Hospital, 1 Blackfan Circle, Room 6006G, Boston, MA, 02115, USA

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  1. Department of Physiology and Biophysics, Boston University School of Medicine, Boston, MA, 02118, USA

    Sunita R. Patel & Judith D. Saide

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Correspondence to Judith D. Saide.

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Patel, S.R., Saide, J.D. Stretchin-klp, a novel Drosophila indirect flight muscle protein, has both myosin dependent and independent isoforms. J Muscle Res Cell Motil 26, 213–224 (2005). https://doi.org/10.1007/s10974-005-9012-y

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