Bruno K S, Aramayo R, Minke P F, Metzenberg R L, Plamann M
Department of Biology, Texas A & M University, College Station 77843-3258, USA.
EMBO J. 1996 Nov 1;15(21):5772-82.
In filamentous fungi, growth polarity (i.e. hyphal extension) and formation of septa require polarized deposition of new cell wall material. To explore this process, we analyzed a conditional Neurospora crassa mutant, mcb, which showed a complete loss of growth polarity when incubated at the restrictive temperature. Cloning and DNA sequence analysis of the mcb gene revealed that it encodes a regulatory subunit of cAMP-dependent protein kinase (PKA). Unexpectedly, the mcb mutant still formed septa when grown at the restrictive temperature, indicating that polarized deposition of wall material during septation is a process that is, at least in part, independent of polarized deposition during hyphal tip extension. However, septa formed in the mcb mutant growing at the restrictive temperature are mislocalized. Both polarized growth and septation are actin-dependent processes, and a concentration of actin patches is observed at growing hyphal tips and sites where septa are being formed. In the mcb mutant growing at the restrictive temperature, actin patches are uniformly distributed over the cell cortex; however, actin patches are still concentrated at sites of septation. Our results suggest that the PKA pathway regulates hyphal growth polarity, possibly through organizing actin patches at the cell cortex.
在丝状真菌中,生长极性(即菌丝延伸)和隔膜的形成需要新细胞壁物质的极化沉积。为了探究这一过程,我们分析了一种条件性粗糙脉孢菌突变体mcb,该突变体在限制温度下培养时表现出完全丧失生长极性。对mcb基因的克隆和DNA序列分析表明,它编码一种依赖于cAMP的蛋白激酶(PKA)的调节亚基。出乎意料的是,mcb突变体在限制温度下生长时仍能形成隔膜,这表明隔膜形成过程中壁物质的极化沉积至少部分独立于菌丝顶端延伸过程中的极化沉积。然而,在限制温度下生长的mcb突变体中形成的隔膜位置错误。极化生长和隔膜形成都是依赖于肌动蛋白的过程,并且在生长的菌丝顶端和正在形成隔膜的部位观察到肌动蛋白斑的聚集。在限制温度下生长的mcb突变体中,肌动蛋白斑均匀地分布在细胞皮层上;然而,肌动蛋白斑仍集中在隔膜形成部位。我们的结果表明,PKA途径可能通过在细胞皮层组织肌动蛋白斑来调节菌丝生长极性。