The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, China.
Gene. 2013 Dec 10;532(1):53-62. doi: 10.1016/j.gene.2013.09.008. Epub 2013 Sep 12.
In this study, we investigated the gene sequence and characteristic of kifc1 in Sepiella maindroni through PCR and RACE technology. Our research aimed particularly at the spatio-temporal expression pattern of kifc1 in the developmental testis through in situ hybridization. The particular role of kifc1 in the spermatogenesis of S. maindroni was our particular interest. Based on multiple protein sequence alignments of KIFC1 homologues, kifc1 gene from the testis of S. maindroni was identified, which consisted of 2432bp including a 2109 in-frame ORF corresponding to 703 continuous amino acids. The encoded polypeptide shared highest similarity with Octopus tankahkeei. Through the prediction of the secondary and tertiary structures, the motor domain of KIFC1 was conserved at the C-terminal, having putative ATP-binding and microtubule-binding motifs, while the N-terminal was more specific to bind various cargoes for cellular events. The stalk domain connecting between the C-terminal and N-terminal determined the direction of movement. According to RT-PCR results, the kifc1 gene is not tissue-specific, commonly detected in different tissues, for example, the testis, liver, stomach, muscle, caecum and gills. Through an in situ hybridization method, the expression pattern of KIFC1 protein mimics in the spermatogenesis of S. maindroni. During the primary stage of the spermatogenesis, the kifc1 mRNA signal was barely detectable. At the early spermatids, the signal started to be present. With the elongation of spermatids, the signals increased substantially. It peaked and gathered around the acrosome area when the spermatids began to transform to spindle shape. As the spermatids developed into mature sperm, the signal vanished. In summary, the expression of kfic1 at specific stages during spermiogenesis and its distribution shed light on the potential functions of this motor in major cytological transformations. The KIFC1 homologue may provide a direct shaping force to the nucleus or influence the shaping process through indirect regulation.
在这项研究中，我们通过 PCR 和 RACE 技术研究了鱿鱼 kifc1 的基因序列和特征。我们的研究特别关注 kifc1 在发育中的睾丸中的时空表达模式，通过原位杂交进行研究。kifc1 在鱿鱼精子发生中的特定作用是我们特别感兴趣的。基于 KIFC1 同源物的多个蛋白质序列比对，鉴定了来自鱿鱼睾丸的 kifc1 基因，该基因由 2432bp 组成，包括 2109 个框内 ORF，对应 703 个连续氨基酸。编码的多肽与章鱼的相似度最高。通过二级和三级结构的预测，KIFC1 的马达结构域在 C 端保守，具有潜在的 ATP 结合和微管结合基序，而 N 端更特异性地结合各种货物以进行细胞事件。连接 C 端和 N 端的茎结构域决定了运动的方向。根据 RT-PCR 结果，kifc1 基因不是组织特异性的，在不同组织中普遍检测到，例如睾丸、肝脏、胃、肌肉、盲肠和鳃。通过原位杂交方法，研究了 KIFC1 蛋白在鱿鱼精子发生中的表达模式。在精子发生的初级阶段，kifc1 mRNA 信号几乎无法检测到。在早期精子中，信号开始出现。随着精子的伸长，信号显著增加。当精子开始转化为纺锤形时，信号达到峰值并聚集在顶体区域周围。随着精子发育成成熟精子，信号消失。总之，在精子发生的特定阶段 kifc1 的表达及其分布揭示了该马达在主要细胞学转化中的潜在功能。KIFC1 同源物可能为核提供直接的成形力，或通过间接调节影响成形过程。
