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Cloning and characterization of the altA -tubulin gene of Physarum

¹McArdle Laboratory for Cancer Research, University of Wisconsin-Madison, 1400 University Avenue, Madison, WI 53706, USA
²Department of Biology, Colgate University, Hamilton, NY 13346, USA
³ Department of Physiology, 719 Veterinary Research Tower, Cornell University, Ithaca, NY 14853–6401, USA

^* Author for correspondence. Tel. (608) 262 0982; fax (608) 262 2824.

ABSTRACT

A cDNA clone derived from the altA locus, encoding one of several -tubulins in Physarum, was sequenced and used to determine the developmental and cell cycle expression patterns of its corresponding gene. The predicted amino acid sequence of the altA gene product, 1A-tubulin, is 92% identical to the other known Physarum -tubulins, 1B and 2B, which are products of two tightly linked genes at the altB locus. The nucleotide sequence of the altA coding region is 82% identical to the two altB genes. Expression of the altA gene was found in all three cell types examined — amoeba, flagellate and plasmodium — but at substantially different levels in each. The peak level of altA message detected in flagellates was 14-fold higher than in amoebae, while the peak level in plasmodia was 5-fold lower than in amoebae. The expression pattern of altA and the predicted amino acid sequence of the -tubulin it encodes suggest that 1A is the substrate for post-translational acetylation, giving rise to the 3-tubulin isoform found specifically in amoebae and flagellates. Northern blot analysis of plasmodial RNA samples from specific times in the cell cycle showed that the level of altA message varies over the cell cycle in a pattern similar to transcripts from other tubulin genes, with a peak at mitosis and little or no message detected during most of interphase.

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