Myosin rod protein: a novel thick filament component of Drosophila muscle.
Myosin rod protein (MRP), a 155 kDa protein encoded by a gene internal to the Drosophila muscle myosin heavy chain (Mhc) gene, contains the MHC rod domain, but has 77 unique N-terminal residues that exactly replace the MHC motor and light chain binding domains. Originally described as an abundant testis protein, we now demonstrate the MRP also is a major component of myofilaments in Drosophila. Specifically, the Mrp promoter directs the expression of a LacZ reporter transgene in somatic, cardiac and visceral muscles. MRP-specific antibodies detect the protein in detergent-insoluble fractions of muscle extracts and co-localize the protein with MHC to the sarcomeric A-band in immunostained muscles. Immunoblot analysis shows that in a set of adult direct flight muscles (DFM), the ratio of MRP to MHC is 1:3. Chemical cross-link and co-immunoprecipitation experiments using 0.5 M KCl-extracted thick filament proteins indicate that native MRP is a homodimer. Electron microscopy of DFM49, which has a high MRP content, shows in cross section, disordered myofilament packing and a variable thin to thick filament ratio and, in longitudinal section, severely bent thin filaments that are not well associated with thick filaments. In rigor, thick filaments from DFM49 consist of segments with cross bridges that are interspersed with smooth domains lacking cross bridges. These data indicate that MRP is a novel contractile protein that co-integrates with myosin into the thick filament, thereby changing structure and function of the sarcomere.
Standiford
D M
DM
Department of Cell and Developmental Biology and Pennsylvania Muscle Institute, University of Pennsylvania School of Medicine, Philadelphia 19104-6058, USA.
Davis
M B
MB
Miedema
K
K
Franzini-Armstrong
C
C
Emerson
C P
CP
D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D004330:Drosophila / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D005260:Female; D005786:Gene Expression Regulation; D017344:Genes, Insect; D008297:Male; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009132:Muscles / Q000378:metabolism / Q000648:ultrastructure; D018995:Myosin Heavy Chains / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D012518:Sarcomeres / Q000378:metabolism / Q000648:ultrastructure; D017386:Sequence Homology, Amino Acid
D004247:DNA; D018995:Myosin Heavy Chains