Two microtubule-plus-end binding proteins LIS1-1 and LIS1-2, homologues of human LIS1 in Neurospora crassa

Olga A. Callejas-Negrete, Michael Plamann, Robert Schnittker, Salomon Bartnicki-García, Robert Roberson, Genaro Pimienta, Rosa R. Mouriño-Pérez

Research output: Contribution to journalArticle

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Abstract

LIS1 is a microtubule (Mt) plus-end binding protein that interacts with the dynein/dynactin complex. In humans, LIS1 is required for proper nuclear and organelle migration during cell growth. Although gene duplication is absent from Neurospora crassa, we found two paralogues of human LIS1. We named them LIS1-1 and LIS1-2 and studied their dynamics and function by fluorescent tagging. At the protein level, LIS1-1 and LIS1-2 were very similar. Although, the characteristic coiled-coil motif was not present in LIS1-2. LIS1-1-GFP and LIS1-2-GFP showed the same cellular distribution and dynamics, but LIS1-2-GFP was less abundant. Both LIS1 proteins were found in the subapical region as single fluorescent particles traveling toward the cell apex, they accumulated in the apical dome forming prominent short filament-like structures, some of which traversed the Spitzenkörper (Spk). The fluorescent structures moved exclusively in anterograde fashion along straight paths suggesting they traveled on Mts. There was no effect in the filament behavior of LIS1-1-GFP in the δ. lis1-. 2 mutant but the dynamics of LIS1-2-GFP was affected in the δ. lis1-. 1 mutant. Microtubular integrity and the dynein-dynactin complex were necessary for the formation of filament-like structures of LIS1-1-GFP in the subapical and apical regions; however, conventional kinesin (KIN-1) was not. Deletion mutants showed that the lack of lis1-. 1 decreased cell growth by ~75%; however, the lack of lis1-. 2 had no effect on growth. A δ. lis1-. 1;δ. lis1-. 2 double mutant showed slower growth than either single mutant. Conidia production was reduced but branching rate increased in δ. lis1-. 1 and the δ. lis1-. 1;δ. lis1-. 2 double mutants. The absence of LIS1-1 had a strong effect on Mt organization and dynamics and indirectly affected nuclear and mitochondrial distribution. The absence of LIS1-1 filaments in dynein mutants (ropy mutants) or in benomyl treated hyphae indicates the strong association between this protein and the regulation of the dynein-dynactin complex and Mt organization. LIS1-1 and LIS1-2 had a high amino acid homology, nevertheless, the absence of the coiled-coil motif in LIS1-2 suggests that its function or regulation may be distinct from that of LIS1-1.

Original languageEnglish (US)
Pages (from-to)213-227
Number of pages15
JournalFungal Genetics and Biology
Volume82
DOIs
StatePublished - Sep 1 2015

Fingerprint

Dyneins
Neurospora crassa
Microtubules
Carrier Proteins
Growth
Benomyl
Kinesin
Proteins
Gene Duplication
Fungal Spores
Hyphae
Organelles
Cell Movement
Amino Acids
Dynactin Complex

Keywords

  • +TIP proteins
  • Dynein-dynactin
  • LIS1
  • Microtubules
  • Neurospora crassa

ASJC Scopus subject areas

  • Genetics
  • Microbiology

Cite this

Callejas-Negrete, O. A., Plamann, M., Schnittker, R., Bartnicki-García, S., Roberson, R., Pimienta, G., & Mouriño-Pérez, R. R. (2015). Two microtubule-plus-end binding proteins LIS1-1 and LIS1-2, homologues of human LIS1 in Neurospora crassa. Fungal Genetics and Biology, 82, 213-227. https://doi.org/10.1016/j.fgb.2015.07.009

Two microtubule-plus-end binding proteins LIS1-1 and LIS1-2, homologues of human LIS1 in Neurospora crassa. / Callejas-Negrete, Olga A.; Plamann, Michael; Schnittker, Robert; Bartnicki-García, Salomon; Roberson, Robert; Pimienta, Genaro; Mouriño-Pérez, Rosa R.

In: Fungal Genetics and Biology, Vol. 82, 01.09.2015, p. 213-227.

Research output: Contribution to journalArticle

Callejas-Negrete, OA, Plamann, M, Schnittker, R, Bartnicki-García, S, Roberson, R, Pimienta, G & Mouriño-Pérez, RR 2015, 'Two microtubule-plus-end binding proteins LIS1-1 and LIS1-2, homologues of human LIS1 in Neurospora crassa', Fungal Genetics and Biology, vol. 82, pp. 213-227. https://doi.org/10.1016/j.fgb.2015.07.009
Callejas-Negrete, Olga A. ; Plamann, Michael ; Schnittker, Robert ; Bartnicki-García, Salomon ; Roberson, Robert ; Pimienta, Genaro ; Mouriño-Pérez, Rosa R. / Two microtubule-plus-end binding proteins LIS1-1 and LIS1-2, homologues of human LIS1 in Neurospora crassa. In: Fungal Genetics and Biology. 2015 ; Vol. 82. pp. 213-227.
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abstract = "LIS1 is a microtubule (Mt) plus-end binding protein that interacts with the dynein/dynactin complex. In humans, LIS1 is required for proper nuclear and organelle migration during cell growth. Although gene duplication is absent from Neurospora crassa, we found two paralogues of human LIS1. We named them LIS1-1 and LIS1-2 and studied their dynamics and function by fluorescent tagging. At the protein level, LIS1-1 and LIS1-2 were very similar. Although, the characteristic coiled-coil motif was not present in LIS1-2. LIS1-1-GFP and LIS1-2-GFP showed the same cellular distribution and dynamics, but LIS1-2-GFP was less abundant. Both LIS1 proteins were found in the subapical region as single fluorescent particles traveling toward the cell apex, they accumulated in the apical dome forming prominent short filament-like structures, some of which traversed the Spitzenk{\"o}rper (Spk). The fluorescent structures moved exclusively in anterograde fashion along straight paths suggesting they traveled on Mts. There was no effect in the filament behavior of LIS1-1-GFP in the δ. lis1-. 2 mutant but the dynamics of LIS1-2-GFP was affected in the δ. lis1-. 1 mutant. Microtubular integrity and the dynein-dynactin complex were necessary for the formation of filament-like structures of LIS1-1-GFP in the subapical and apical regions; however, conventional kinesin (KIN-1) was not. Deletion mutants showed that the lack of lis1-. 1 decreased cell growth by ~75{\%}; however, the lack of lis1-. 2 had no effect on growth. A δ. lis1-. 1;δ. lis1-. 2 double mutant showed slower growth than either single mutant. Conidia production was reduced but branching rate increased in δ. lis1-. 1 and the δ. lis1-. 1;δ. lis1-. 2 double mutants. The absence of LIS1-1 had a strong effect on Mt organization and dynamics and indirectly affected nuclear and mitochondrial distribution. The absence of LIS1-1 filaments in dynein mutants (ropy mutants) or in benomyl treated hyphae indicates the strong association between this protein and the regulation of the dynein-dynactin complex and Mt organization. LIS1-1 and LIS1-2 had a high amino acid homology, nevertheless, the absence of the coiled-coil motif in LIS1-2 suggests that its function or regulation may be distinct from that of LIS1-1.",
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T1 - Two microtubule-plus-end binding proteins LIS1-1 and LIS1-2, homologues of human LIS1 in Neurospora crassa

AU - Callejas-Negrete, Olga A.

AU - Plamann, Michael

AU - Schnittker, Robert

AU - Bartnicki-García, Salomon

AU - Roberson, Robert

AU - Pimienta, Genaro

AU - Mouriño-Pérez, Rosa R.

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N2 - LIS1 is a microtubule (Mt) plus-end binding protein that interacts with the dynein/dynactin complex. In humans, LIS1 is required for proper nuclear and organelle migration during cell growth. Although gene duplication is absent from Neurospora crassa, we found two paralogues of human LIS1. We named them LIS1-1 and LIS1-2 and studied their dynamics and function by fluorescent tagging. At the protein level, LIS1-1 and LIS1-2 were very similar. Although, the characteristic coiled-coil motif was not present in LIS1-2. LIS1-1-GFP and LIS1-2-GFP showed the same cellular distribution and dynamics, but LIS1-2-GFP was less abundant. Both LIS1 proteins were found in the subapical region as single fluorescent particles traveling toward the cell apex, they accumulated in the apical dome forming prominent short filament-like structures, some of which traversed the Spitzenkörper (Spk). The fluorescent structures moved exclusively in anterograde fashion along straight paths suggesting they traveled on Mts. There was no effect in the filament behavior of LIS1-1-GFP in the δ. lis1-. 2 mutant but the dynamics of LIS1-2-GFP was affected in the δ. lis1-. 1 mutant. Microtubular integrity and the dynein-dynactin complex were necessary for the formation of filament-like structures of LIS1-1-GFP in the subapical and apical regions; however, conventional kinesin (KIN-1) was not. Deletion mutants showed that the lack of lis1-. 1 decreased cell growth by ~75%; however, the lack of lis1-. 2 had no effect on growth. A δ. lis1-. 1;δ. lis1-. 2 double mutant showed slower growth than either single mutant. Conidia production was reduced but branching rate increased in δ. lis1-. 1 and the δ. lis1-. 1;δ. lis1-. 2 double mutants. The absence of LIS1-1 had a strong effect on Mt organization and dynamics and indirectly affected nuclear and mitochondrial distribution. The absence of LIS1-1 filaments in dynein mutants (ropy mutants) or in benomyl treated hyphae indicates the strong association between this protein and the regulation of the dynein-dynactin complex and Mt organization. LIS1-1 and LIS1-2 had a high amino acid homology, nevertheless, the absence of the coiled-coil motif in LIS1-2 suggests that its function or regulation may be distinct from that of LIS1-1.

AB - LIS1 is a microtubule (Mt) plus-end binding protein that interacts with the dynein/dynactin complex. In humans, LIS1 is required for proper nuclear and organelle migration during cell growth. Although gene duplication is absent from Neurospora crassa, we found two paralogues of human LIS1. We named them LIS1-1 and LIS1-2 and studied their dynamics and function by fluorescent tagging. At the protein level, LIS1-1 and LIS1-2 were very similar. Although, the characteristic coiled-coil motif was not present in LIS1-2. LIS1-1-GFP and LIS1-2-GFP showed the same cellular distribution and dynamics, but LIS1-2-GFP was less abundant. Both LIS1 proteins were found in the subapical region as single fluorescent particles traveling toward the cell apex, they accumulated in the apical dome forming prominent short filament-like structures, some of which traversed the Spitzenkörper (Spk). The fluorescent structures moved exclusively in anterograde fashion along straight paths suggesting they traveled on Mts. There was no effect in the filament behavior of LIS1-1-GFP in the δ. lis1-. 2 mutant but the dynamics of LIS1-2-GFP was affected in the δ. lis1-. 1 mutant. Microtubular integrity and the dynein-dynactin complex were necessary for the formation of filament-like structures of LIS1-1-GFP in the subapical and apical regions; however, conventional kinesin (KIN-1) was not. Deletion mutants showed that the lack of lis1-. 1 decreased cell growth by ~75%; however, the lack of lis1-. 2 had no effect on growth. A δ. lis1-. 1;δ. lis1-. 2 double mutant showed slower growth than either single mutant. Conidia production was reduced but branching rate increased in δ. lis1-. 1 and the δ. lis1-. 1;δ. lis1-. 2 double mutants. The absence of LIS1-1 had a strong effect on Mt organization and dynamics and indirectly affected nuclear and mitochondrial distribution. The absence of LIS1-1 filaments in dynein mutants (ropy mutants) or in benomyl treated hyphae indicates the strong association between this protein and the regulation of the dynein-dynactin complex and Mt organization. LIS1-1 and LIS1-2 had a high amino acid homology, nevertheless, the absence of the coiled-coil motif in LIS1-2 suggests that its function or regulation may be distinct from that of LIS1-1.

KW - +TIP proteins

KW - Dynein-dynactin

KW - LIS1

KW - Microtubules

KW - Neurospora crassa

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