Kinesin 6 regulation in Drosophila female meiosis by the non-conserved N- and C- terminal domains

Arunika Das, Jeffry Cesario, Anna Maria Hinman, Janet K. Jang, Kim McKim

Research output: Contribution to journalArticle

Abstract

Bipolar spindle assembly occurs in the absence of centrosomes in the oocytes of most organisms. In the absence of centrosomes in Drosophila oocytes, we have proposed that the kinesin 6 Subito, a MKLP-2 homolog, is required for establishing spindle bipolarity and chromosome biorientation by assembling a robust central spindle during prometaphase I. Although the functions of the conserved motor domains of kinesins is well studied, less is known about the contribution of the poorly conserved N- and C- terminal domains to motor function. In this study, we have investigated the contribution of these domains to kinesin 6 functions in meiosis and early embryonic development. We found that the N-terminal domain has antagonistic elements that regulate localization of the motor to microtubules. Other parts of the N- and C-terminal domains are not required for microtubule localization but are required for motor function. Some of these elements of Subito are more important for either mitosis or meiosis, as revealed by separation-of-function mutants. One of the functions for both the N- and C-terminals domains is to restrict the CPC to the central spindle in a ring around the chromosomes. We also provide evidence that CDK1 phosphorylation of Subito regulates its activity associated with homolog bi-orientation. These results suggest the N- and C-terminal domains of Subito, while not required for localization to the central spindle microtubules, have important roles regulating Subito, by interacting with other spindle proteins and promoting activities such as bipolar spindle formation and homologous chromosome bi-orientation during meiosis.

Original languageEnglish (US)
Pages (from-to)1555-1569
Number of pages15
JournalG3: Genes, Genomes, Genetics
Volume8
Issue number5
DOIs
StatePublished - May 1 2018

Fingerprint

Kinesin
Meiosis
Microtubules
Drosophila
Centrosome
Oocytes
Chromosomes
Prometaphase
Ring Chromosomes
Mitosis
Embryonic Development
Phosphorylation
Proteins

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics
  • Genetics(clinical)

Keywords

  • Chromosome segregation
  • Drosophila
  • Kinesin 6
  • Meiosis
  • Microtubule
  • Oocyte

Cite this

Das, Arunika ; Cesario, Jeffry ; Hinman, Anna Maria ; Jang, Janet K. ; McKim, Kim. / Kinesin 6 regulation in Drosophila female meiosis by the non-conserved N- and C- terminal domains. In: G3: Genes, Genomes, Genetics. 2018 ; Vol. 8, No. 5. pp. 1555-1569.
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Kinesin 6 regulation in Drosophila female meiosis by the non-conserved N- and C- terminal domains. / Das, Arunika; Cesario, Jeffry; Hinman, Anna Maria; Jang, Janet K.; McKim, Kim.

In: G3: Genes, Genomes, Genetics, Vol. 8, No. 5, 01.05.2018, p. 1555-1569.

Research output: Contribution to journalArticle

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T1 - Kinesin 6 regulation in Drosophila female meiosis by the non-conserved N- and C- terminal domains

AU - Das, Arunika

AU - Cesario, Jeffry

AU - Hinman, Anna Maria

AU - Jang, Janet K.

AU - McKim, Kim

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N2 - Bipolar spindle assembly occurs in the absence of centrosomes in the oocytes of most organisms. In the absence of centrosomes in Drosophila oocytes, we have proposed that the kinesin 6 Subito, a MKLP-2 homolog, is required for establishing spindle bipolarity and chromosome biorientation by assembling a robust central spindle during prometaphase I. Although the functions of the conserved motor domains of kinesins is well studied, less is known about the contribution of the poorly conserved N- and C- terminal domains to motor function. In this study, we have investigated the contribution of these domains to kinesin 6 functions in meiosis and early embryonic development. We found that the N-terminal domain has antagonistic elements that regulate localization of the motor to microtubules. Other parts of the N- and C-terminal domains are not required for microtubule localization but are required for motor function. Some of these elements of Subito are more important for either mitosis or meiosis, as revealed by separation-of-function mutants. One of the functions for both the N- and C-terminals domains is to restrict the CPC to the central spindle in a ring around the chromosomes. We also provide evidence that CDK1 phosphorylation of Subito regulates its activity associated with homolog bi-orientation. These results suggest the N- and C-terminal domains of Subito, while not required for localization to the central spindle microtubules, have important roles regulating Subito, by interacting with other spindle proteins and promoting activities such as bipolar spindle formation and homologous chromosome bi-orientation during meiosis.

AB - Bipolar spindle assembly occurs in the absence of centrosomes in the oocytes of most organisms. In the absence of centrosomes in Drosophila oocytes, we have proposed that the kinesin 6 Subito, a MKLP-2 homolog, is required for establishing spindle bipolarity and chromosome biorientation by assembling a robust central spindle during prometaphase I. Although the functions of the conserved motor domains of kinesins is well studied, less is known about the contribution of the poorly conserved N- and C- terminal domains to motor function. In this study, we have investigated the contribution of these domains to kinesin 6 functions in meiosis and early embryonic development. We found that the N-terminal domain has antagonistic elements that regulate localization of the motor to microtubules. Other parts of the N- and C-terminal domains are not required for microtubule localization but are required for motor function. Some of these elements of Subito are more important for either mitosis or meiosis, as revealed by separation-of-function mutants. One of the functions for both the N- and C-terminals domains is to restrict the CPC to the central spindle in a ring around the chromosomes. We also provide evidence that CDK1 phosphorylation of Subito regulates its activity associated with homolog bi-orientation. These results suggest the N- and C-terminal domains of Subito, while not required for localization to the central spindle microtubules, have important roles regulating Subito, by interacting with other spindle proteins and promoting activities such as bipolar spindle formation and homologous chromosome bi-orientation during meiosis.

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