A temperature-sensitive mutation of the Schizosaccharomyces pombe gene nuc2+ that encodes a nuclear scaffold-like protein blocks spindle elongation in mitotic anaphase.
A temperature-sensitive mutation of the Schizosaccharomyces pombe gene nuc2+ that encodes a nuclear scaffold-like protein blocks spindle elongation in mitotic anaphase.
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DOI：
10.1083/jcb.106.4.1171
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发表时间：
1988-04
期刊：
影响因子：
--
通讯作者：
Yanagida M
中科院分区：
文献类型：
--
作者：
Hirano T;Hiraoka Y;Yanagida M
裂殖酵母裂殖酵母的温度敏感突变体nuc 2 -663在限制性温度下特异性地阻断有丝分裂纺锤体的伸长，使得被捕细胞中的细胞核包含短的均匀纺锤体（约3微米长），其贯穿由三个浓缩染色体组成的中期板状结构。在野生型或突变体细胞中，在允许温度下，纺锤体在后期完全延伸约15微米长。通过转化将nuc 2 '基因克隆到2.4kb的基因组DNA片段中，并测定了其完整的核苷酸序列。它的编码区预测665个残基的内部重复蛋白质（76.250摩尔重量）。通过使用抗lacZ-nuc 2+融合蛋白的抗血清进行免疫印迹，在野生型S中检测到由nuc 2+编码的多肽（命名为p67; 67，000 mol wt）。p67的表达量在将携带nuc 2+基因的多拷贝或高表达质粒导入S.粟酒裂殖酵母细胞细胞分级分离和Percoll梯度离心结合免疫印迹显示，p67与细胞核共分级分离，并富含耐药结构，该耐药结构不溶于2 M NaCl、25 mM 3，5 '-二碘水杨酸锂和1% Triton，但可溶于8 M尿素。然而，在nuc 2突变细胞中，可溶性p76，可能是一种未加工的前体，除了不溶性p67外，也积累了。nuc 2+基因的作用可能是在染色体分离中连接核和细胞骨架功能。
A temperature-sensitive mutant nuc2-663 of the fission yeast Schizosaccharomyces pombe specifically blocks mitotic spindle elongation at restrictive temperature so that nuclei in arrested cells contain a short uniform spindle (approximately 3-micron long), which runs through a metaphase plate-like structure consisting of three condensed chromosomes. In the wild-type or in the mutant cells at permissive temperature, the spindle is fully extended approximately 15- micron long in anaphase. The nuc2' gene was cloned in a 2.4-kb genomic DNA fragment by transformation, and its complete nucleotide sequence was determined. Its coding region predicts a 665-residues internally repeating protein (76.250 mol wt). By immunoblots using anti-sera raised against lacZ-nuc2+ fused proteins, a polypeptide (designated p67; 67,000 mol wt) encoded by nuc2+ is detected in the wild-type S. pombe extracts; the amount of p67 is greatly increased when multi-copy or high-expression plasmids carrying the nuc2+ gene are introduced into the S. pombe cells. Cellular fractionation and Percoll gradient centrifugation combined with immunoblotting show that p67 cofractionates with nuclei and is enriched in resistant structure that is insoluble in 2 M NaCl, 25 mM lithium 3,5'-diiodosalicylate, and 1% Triton but is soluble in 8 M urea. In nuc2 mutant cells, however, soluble p76, perhaps an unprocessed precursor, accumulates in addition to insoluble p67. The role of nuc2+ gene may be to interconnect nuclear and cytoskeletal functions in chromosome separation.
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DOI：
10.1083/jcb.98.1.352
发表时间：
1984-01
期刊：
The Journal of cell biology
影响因子：
--
作者：
Earnshaw WC;Halligan N;Cooke C;Rothfield N
通讯作者：
Rothfield N
影响因子：
3.2
作者：
ITO, H;FUKUDA, Y;KIMURA, A
通讯作者：
KIMURA, A
影响因子：
3.7
作者：
COMINGS, DE;OKADA, TA
通讯作者：
OKADA, TA
影响因子：
11.4
作者：
HIRANO, T;FUNAHASHI, S;YANAGIDA, M
通讯作者：
YANAGIDA, M
DOI：
10.1073/pnas.72.3.1007
发表时间：
1975-01-01
影响因子：
11.1
作者：
AARONSON, RP;BLOBEL, G
通讯作者：
BLOBEL, G