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Molecular Biology and Gene Regulation

T-URF 13 Protein from Mitochondria of Texas Male-Sterile Maize (Zea mays L.) ¹

Its Purification and Submitochondrial Localization, and Immunogold Labeling in Anther Tapetum during Microsporogenesis

Department of Botany, Iowa State University, Ames, Iowa 50011-1020

The protein T-URF13 (URF13) is specific to mitochondria of maize (Zea mays L.) with Texas (T) male-sterile cytoplasm and has been implicated in causing male sterility and susceptibility to T-cytoplasm-specific fungal diseases. T-URF13 was purified from isolated mitochondria from maize (line B73) with T cytoplasm by gel filtration and a quasi two-dimensional polyacrylamide gel electrophoresis system. Antibodies to the purified and denatured protein were produced in rabbits. Anti-T-URF13 antiserum was used to show that T-URF13 is in the inner membrane of mitochondria and behaves as an integral membrane protein when mitochondria are fractionated with sodium carbonate or Triton X-114. The antiserum and protein A tagged with 20-nanometer-gold particles were used to localize T-URF13 in T mitochondria by electron microscopy of sections of isolated mitochondria from etiolated shoots and sections of roots and of tapetal cells at pre-and post-degeneration stages of microsporogenesis. The microscopic study confirms that T-URF13 is specifically localized in the mitochondrial membranes of all of the T mitochondria tested, notably those in the tapetum from the meiocyte stage to the late-microspore stage. No change in the amount of labeled T-URF13 protein in the mitochondria of aging tapetal cells was detected.

³ Present address: Genetics Program, Michigan State University, East Lansing, MI 48824.

¹ Supported by National Science Foundation grant DCB-8409797, by a grant from Enichem Americas, Inc., Biotechnology Laboratory, and by the Department of Botany and the Graduate College, Iowa State University, Ames.

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