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MEMBRANE TRANSPORTERS, ION CHANNELS, AND PUMPS

Accumulation of _m, a structural member of X,K-ATPase -subunit family, in nuclear envelopes of perinatal myocytes

¹Department of Pharmacology, Medical College of Ohio, Toledo, Ohio 43614; and ²Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow 117997, Russia

Submitted 22 August 2003 ; accepted in final form 26 November 2003

Recently discovered muscle-specific _m protein is structurally closely related to the X,K-ATPase -subunits. However, it has a number of unique properties such as predominant localization in intracellular stores and lack of association with known X,K-ATPase -subunits on heterologous coexpression. In this study, the primary structure of mouse _m was determined and developmental regulation of the gene (ATP1B4) was analyzed. The expression is first detected at day 14 of gestation, is sharply increased at day 16, and reaches its maximum at day 18. After birth, the expression quickly decreases and is hardly detectable in adult mice. A more detailed subcellular localization study was undertaken, and its results indicate that _m not only is located in sarcoplasmic reticulum but is concentrated in nuclear envelopes of both prenatal and postnatal skeletal muscles. Immunohistochemical studies show that _m is specific to myocytes and, at the subcellular level, many nuclear envelopes are intensively labeled in both fetal and newborn skeletal muscles. Accordingly, _m is detected by immunoblotting in purified nuclei and nuclear membranes from neonatal skeletal muscles. On transfection of human rhabdomyosarcoma cell line RD, green fluorescent protein-tagged _m resides intracellularly with significant enrichment in nuclear envelopes, whereas _m with transmembrane domain deleted localizes in both cytoplasm and nucleoplasm. Nuclear _m apparently is not in association with Na,K-ATPase because we never detected its -subunit in myonuclear membranes. These results indicate that _m has a specialized function in mammalian perinatal myocytes, different from functions of other X,K-ATPase -subunits. The unique temporospatial distribution of _m protein expression suggests its important role in development of growing skeletal muscle.

ATP1B4; sodium, potassium-adenosine 5'-triphosphatase; nuclear membrane; skeletal muscle development

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