DOI: 10.5061/dryad.mcvdnck8n
title: Blocking microtubule deacetylation inhibits anaphase chromosome movements in crane-fly spermatocytes

description: Chromosome movement speeds during anaphase are regulated by
 depolymerization of microtubules. Several models describe chromosome
 movement during cell division but none of them consider post-translational
 modifications of tubulin, even though such modifications help specify
 microtubules for unique cellular activities. Among these modifications,
 acetylation of Lysine 40 is one of the common post-translational
 modifications. Acetylation of microtubules greatly improves their
 stability, especially when subjected to cooling or drug treatment. Since
 kinetochore microtubules are acetylated in a variety of eukaryote cells,
 we wondered whether deacetylation of kinetochore microtubules was
 necessary in order for microtubules to be able to depolymerize during
 anaphase. HDAC6 (Histone Deacetylase 6) deacetylates acetylated tubulin.
 To study whether tubulin must be deacetylated during anaphase, we added to
 living cells two different HDAC6 inhibitors (Tubacin and Trichostatin A),
 separately, as chromosomes moved poleward in anaphase. Both HDAC6
 inhibitors altered chromosome movement: chromosomes either completely
 stopped moving, or moved more slowly, or sometimes continued movement
 without speed changes. The effects of the inhibitors on chromosome
 movement are reversible: half-bivalents either restarted anaphase movement
 by themselves or resumed their poleward movement after the inhibitor was
 washed out. We suggest that kinetochore microtubules need to be
 deacetylated in order for normal anaphase movements to occur.
description: # Blocking microtubules deacetylation inhibits anaphase chromosome
 movements in crane-fly spermatocytes The data package contains seven
 datasets: 1. Fig_2_.csv 2. Figs_4.csv 3. Fig_5.csv 4.
 Backward_movement.csv 5. Fig_7.csv 6. Table_1_control_cells.csv 7.
 Table_1_treated_cells.csv **(1) Fig_2_.csv** This dataset contains 149N
 values with columns named "Distance (microm)" ,
 "Acetylated_alpha_tubulin", and
 "Tyrosinated_alpha_tubulin". These three columns are the data
 before normalization of the Y axis (gray values of acetylated and
 tyrosinated tubulin) between 0 and 1. The next two columns, named
 "Normalized_Acetylated_alpha_tubulin" and
 "Normalized_Tyrosinated_alpha_tubulin", are the data after
 Y-axis normalization. Variable names with descriptions: * Distance
 (microm): shows the distance from the kinetochore of a microtubule to the
 pole. * Acetylated_alpha_tubulin: shows the gray values (staining
 intensity) of acetylated tubulin of a fix line from the kinetochore to the
 pole. * Tyrosinated_alpha_tubulin: shows the gray value (staining
 intensity) of the same line but on the Tyrosinated tubulin channel. **(2)
 Figs_4.csv** ***Tubacin*** This dataset contains 106N values for 106
 half-bivalents/or 53 pairs of chromosomes, and chromosome speed
 (microm/min) before drug treatment, after drug treatment, and after drug
 washout. Variable names with descriptions: * Drug treatments: shows the
 concentration of Tubacin that was used. * The speed at the beginning
 (microm/min): shows the speed of each half-bivalent from the equatorial
 plane to the poles since the moment that anaphase started up to drug
 perfusion. * The speed after drug treatment (microm/min): shows the speed
 of each half-bivalent since the drug was added. * The speed after drug
 washout (microm/min): shows the speed of each half-bivalent after the drug
 was washed out by Ringer's solution. **(3) Fig_5.csv** ***TSA*** This
 dataset contains 18N values for 18 half-bivalents/ or 9 pairs of
 chromosomes and chromosome speed (microm/min) before drug treatment, after
 drug treatment, and after drug washout. Variable names with descriptions:
 * Drug treatments: shows the concentration of TSA that was used. * The
 speed at the beginning (microm/min): shows the speed of each half-bivalent
 from the equatorial plane to the poles since the moment that anaphase
 started up to drug perfusion. * The speed after drug treatment
 (microm/min): shows the speed of each half-bivalent since the drug was
 added. * The speed after drug washout (microm/min): shows the speed of
 each half-bivalent after the drug was washed out by Ringer's
 solution. **(4) Backward_movement.csv** ***Backward movement*** This
 dataset contains 36N values for 36 half-bivalents/ or 18 pairs of
 chromosomes and the speed of chromosomes (microm/min) moving backward to
 the equatorial plane after drug treatment. Variable names with
 descriptions: * Drug treatments: shows the concentration of Tubacin that
 was used. * Backward movement (microm/min): shows the speed of chromosomes
 moving backward after drug treatment. **(5) Fig_7.csv** This dataset
 contains columns named "Distance (microm)",
 "Control_cell", "TSA-treated", and
 "Tubacin-treated". These four columns are the data before
 normalization of the Y axis (gray values of acetylated tubulin in three
 different cells) between 0 and 1. The next three columns, named
 "Normalized_control_cell", "Normalized_TSA_treated",
 and "Normalized_Tubacin_treated", are the data after Y-axis
 normalization. Variable names with descriptions: * Distance (microm):
 shows the distance from the kinetochore of a microtubule to the pole. *
 Control_cell: shows the gray value or staining intensity of the
 acetylated_tubulin in a normal control cell. * TSA_treated: shows the gray
 value or staining intensity of the acetylated_tubulin in a TSA_treated
 cell. * Tubacin_treated: shows the gray value or staining intensity of the
 acetylated_tubulin in a Tubacin_treated cell. **(6)
 Table_1_control_cells.csv** This dataset contains columns named "KT
 MTs", "FLAGELLUM MTs", "ratio KT MTs/Flagel", and
 "PREDICTED NUMBERS of KT-MTs" in control cells. The cell's
 ID and stage have been added at the top of each table in blue. In this
 dataset, we estimated the number of acetylated microtubules in the
 kinetochore fiber. Variable names with descriptions: * KT MTs: shows the
 maximum intensity of kinetochore microtubules * FLAGELLUM MTs: shows the
 maximum intensity of flagella * ratio KT MTs/Flagel: shows the values from
 dividing the "KT MTs" by "FLAGELLUM MTs". * PREDICTED
 NUMBERS of KT-MTs: shows the values from multiplying the "ratio KT
 MTs/Flagel" by 20, because the flagellum has 20 microtubule
 filaments. **(7) Table_1_treated_cells.csv** This dataset contains columns
 named "KT MTs", "FLAGELLUM MTs", "ratio KT
 MTs/Flagel", and "PREDICTED NUMBERS of KT-MTs" in treated
 cells. The cell's ID and stage have been added at the top of each
 table in blue. In this dataset, we estimated the number of acetylated
 microtubules in the kinetochore fiber. Variable names with descriptions: *
 KT MTs: shows the maximum intensity of kinetochore microtubules *
 FLAGELLUM MTs: shows the maximum intensity of flagella * ratio KT
 MTs/Flagel: shows the values from dividing the "KT MTs" by
 "FLAGELLUM MTs". * PREDICTED NUMBERS of KT-MTs: shows the values
 from multiplying the "ratio KT MTs/Flagel" by 20, because the
 flagellum has 20 microtubule filaments.