tau Regulation of microtubule-microtubule spacing and bundling

Tau proteins are microtubule-associated proteins that promote microtubule polymerization in vitro and in vivo. They are a family of neuronal proteins with apparent molecular weights in the range 50,000-68,000 determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Recently, a new member of this family has been described and its cDNA has been cloned. It has an apparent molecular weight of 116,000 and has been called high-molecular-weight tau (HMW tau). All the tau proteins are encoded by a single gene, which undergoes complex alternative splicing. In the present study, we have cloned into the baculovirus a cDNA fully encoding HMW tau as well as a truncated cDNA encoding a protein beginning 13 amino acids in front of the tau microtubule-binding domain. HMW tau-recombinant-virus-infected Sf9 cells overexpressed HMW tau, which induced the polymerization of microtubules and the formation of long cellular processes similar to those induced by low-molecular-weight tau (LMW tau) overexpression. Process cross sections revealed a larger spacing (approximately 35 nm) between microtubules when induced by HMW tau than when induced by LMW tau (approximately 20 nm). The truncated construct also induces processes, where microtubules were packed far more closely together (approximately 10 nm). Although branching did not occur in processes induced by intact tau S, 10% of the processes induced by the truncated tau protein branched.