Sakakibara H, Takada S, King S M, Witman G B, Kamiya R
Communications Research Laboratory, Kobe, Japan.
J Cell Biol. 1993 Aug;122(3):653-61. doi: 10.1083/jcb.122.3.653.
A new allele of the Chlamydomonas oda4 flagellar mutant (oda4-s7) possessing abnormal outer dynein arms was isolated. Unlike the previously described oda4 axoneme lacking all three (alpha, beta, and gamma) outer-arm dynein heavy chains, the oda4-s7 axoneme contains the alpha and gamma heavy chains and a novel peptide with a molecular mass of approximately 160 kD. The peptide reacts with a mAb (18 beta B) that recognizes an epitope on the NH2-terminal part of the beta heavy chain. These observations indicate that this mutant has a truncated beta heavy chain, and that the NH2-terminal part of the beta heavy chain is important for the stable assembly of the outer arms. In averaged electron microscopic images of outer arms from cross sections of axonemes, the mutant outer arm lacks its mid-portion, producing a forked appearance. Together with our previous finding that the mutant oda11 lacks the alpha heavy chain and the outermost portion of the arm (Sakakibara, H., D. R. Mitchell, and R. Kamiya. 1991. J. Cell Biol. 113:615-622), this result defines the approximate locations of the three outer arm heavy chains in the axonemal cross section. The swimming velocity of oda4-s7 is 65 +/- 8 microns/s, close to that of oda4 which lacks the entire outer arm (62 +/- 8 microns/s) but significantly lower than the velocities of wild type (194 +/- 23 microns/s) and oda11 (119 +/- 17 microns/s). Thus, the lack of the beta heavy chain impairs outer-arm function more seriously than does the lack of the alpha heavy chain, suggesting that the alpha and beta chains play different roles in outer arm function.
分离出一种新的衣藻oda4鞭毛突变体(oda4-s7)的等位基因,其具有异常的外动力蛋白臂。与先前描述的缺乏所有三种(α、β和γ)外臂动力蛋白重链的oda4轴丝不同,oda4-s7轴丝含有α和γ重链以及一种分子量约为160 kD的新肽段。该肽段与一种单克隆抗体(18βB)发生反应,该抗体识别β重链NH2末端部分的一个表位。这些观察结果表明,该突变体具有截短的β重链,并且β重链的NH2末端部分对于外臂的稳定组装很重要。在轴丝横切面外臂的平均电子显微镜图像中,突变体的外臂缺少中部,呈现出叉状外观。连同我们之前的发现,即突变体oda11缺乏α重链和臂的最外部分(Sakakibara, H., D. R. Mitchell, and R. Kamiya. 1991. J. Cell Biol. 113:615 - 622),这一结果确定了轴丝横切面中三种外臂重链的大致位置。oda4-s7的游动速度为65±8微米/秒,接近缺乏整个外臂的oda4的游动速度(62±8微米/秒),但显著低于野生型(194±23微米/秒)和oda11(119±17微米/秒)的速度。因此,β重链的缺失比α重链的缺失更严重地损害外臂功能,这表明α链和β链在外臂功能中发挥不同的作用。