| The dynamic stability of cell dynamics makes the main components and functional components of cells in a dynamic and stable process,maintaining the normal growth and division of cells.The uge1 gene encodes UDP-glucose 4-epimerase 1,which plays an important role in cell growth and maintaining normal cell morphology of sexual reproduction.The rok1 gene predicted to encode an ATP-dependent RNA helicase,which plays an important role in maintaining the normal growth of cells and the normal separation of meiotic chromosomes.At present,there are no reports about the effects of uge1 and rok1 gene deletion on chromosome,spindle and actin dynamics during mitosis.In this project,Schizosaccharomyces pombe was selected as the research object,and the slow-growing strains of uge1Δ and rok1Δ were screened out.By constructing uge1Δ and rok1Δ strains with fluorescent protein markers,the laser confocal live cell imaging was used for the first time to analyze the effects of uge1 and rok1 gene deletion on mitotic chromosome,spindle,and actin dynamics.The results are as follows:(1)The growth results showed that the uge1Δ strain grew extremely significant slower than the wild-type strain in 2-4 h,and after 6 h,the uge1Δstrain grew significantly slower than the wild-type strain.The rok1Δ strain grew extremely significant slower than the wild-type strain after 6 h.The above results suggested that the deletion of uge1 gene had a greater impact on the growth of Schizosaccharomyces pombe.(2)The results of conjugation showed that the number of 4 spores produced by uge1Δ cells was extremely significant reduced compared with that of wild-type cells.Compared with wild-type cells,the number of 4 spores produced by rok1Δ cells was extremely significant reduced,and the number of 3 spores produced by rok1Δ cells was significantly increased.The results showed that the loss of rok1 gene had a greater effect on sporulation.(3)The results of interphase microtubules showed that the deletion of uge1 and rok1 genes led to a extremely significant increase in the number of 5microtubules.Among them,the ratio of 5 microtubules in uge1Δ cells was35.00±5.00%,and the ratio of 5 microtubules in rok1Δ cells was 40.00±5.00%.It was indicating that the deletion of rok1 gene had a greater impact on the number of interphase microtubules.(4)The results of chromosome and spindle dynamics during mitosis showed that the chromosome segregation of uge1Δ cells was abnormal,and unipolar spindle appeared in uge1Δ cells.The time from formation to breakage of the uge1Δ cell spindle was extremely significant increased.The chromosome division of rok1Δ cells was abnormal,and unipolar spindle appeared in rok1Δcells.The time from formation to breakage of the rok1Δ cells spindle was extremely significant increased,and the length of the rok1Δ cells spindle is extremely significant increased.It showed that the deletion of rok1 gene had a more obvious effect on the length of the spindle.Spindle elongation kinetics showed that the deletion of uge1 gene resulted in a significant increase in the prophase elongation rate of the spindle,a extremely significant increase in the prophase time,a extremely significant decrease in the anaphase elongation rate,and a extremely significant increase in the anaphase elongation time.rok1 gene deletion can lead to a extremely significant increase in the prophase elongation time of the spindle,a extremely significant increase in the metaphase elongation time,a extremely significant decrease in the anaphase elongation rate,and a extremely significant increase in the anaphase time.This showed that the deletion of uge1 gene has a significant effect on the elongation rate,and the deletion of rok1 gene had a significant effect on the spindle elongation time.(5)The cell morphology showed that the length of uge1Δ cells was extremely significant increased,the width of uge1Δ cells had no significant difference,and the ratio of length-width of uge1Δ cells was extremely significant increased.The length of rok1Δ cells was extremely significant increased,the width of the rok1Δcells was affected by different degrees,and the ratio of length-width of rok1Δcells also changed accordingly.It showed that the deletion of rok1 gene would cause greater changes in cell morphology.(6)The type of spindle formation showed that the “Bar” spindle of uge1Δcells was extremely significant reduced,and the “Dot” spindle was extremely significant increased.The "Bar" and unipolar spindle of rok1Δ cells were extremely significant reduced,and the " Dot" spindle was significantly increased.It showed that rok1 was more critical to the formation of the spindle.(7)Actin kinetics results showed that the length,the time from polymerization to depolymerization,the residence time and shrinkage time of actin in uge1Δ cells were extremely significant increased,th e rate from polymerization to depolymerization of actin in uge1Δ cells was extremely significant reduced.the length of actin in rok1Δ cells was extremely significant decreased,the time from polymerization to depolymerization of actin in rok1Δcells was significantly increased,the residence time of actin was extremely significant increased,the rate of actin from polymerization to depolymerization was extremely significant decreased.In conclusion,the deletion of uge1 and rok1 genes would affect the dynamics of actin,the deletion of uge1 gene would cause changes in 5 sets of actin dynamics data,and the deletion of rok1 gene would cause changes in 4 sets of actin dynamics data,indicating uge1 gene deletion had a greater impact on the dynamics of fission yeast actin.Severe imbalance of cell dynamics will lead to abnormal cell growth and division,and then lead to related diseases.It was found that the deletion of uge1 gene and rok1 gene would affect the cell growth of Schizosaccharomyces pombe.Moreover,it was found for the first time t hat the deletion of uge1 gene and rok1 gene would affect the mitotic chromosome,spindle and actin dynamics of Schizosaccharomyces pombe.The above research provides a scientific basis for exploring the related dynamic mechanism and related diseases. |
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