1
Supplementary Table 1. List of K. marxianus strains used in this study
Strain name
Km05u
Genotype
a ura3Δ mutant strain derived from K. marxianus
KCTC 17555 (CBS 6556= ATCC 26548)
Reference
(Lee KS et al, 2013)
Km05ku80Δ
ura3Δ ku80::hisG-ScURA3-hisG
this study
Km05ku80Δ-u
ura3Δ ku80::hisG
this study
Km05ku80pdc1Δ
ura3Δ ku80::hisG pdc1:::hisG-ScURA3-hisG
this study
Km05ku80Δpdc5Δ
ura3Δ ku80::hisG pdc5:::hisG-ScURA3-hisG
this study
Km05ku80Δleu2Δ
ura3Δ ku80::hisG leu2:::hisG-ScURA3-hisG
this study
Km05ku80Δleu2Δ-u
ura3Δ ku80::hisG leu2:::hisG
this study
2
3
4
1
5
Supplementary Table 2. List of plasmids used in this study
Plasmid name
pKI
pKI-KU80DsU2
pKI-KmPDC1DU53
pKI-KmPDC5DU2
pKI-KmLEU2DU250
pKI-KmLEU2DU2100
pKI-KmLEU2DU2250
pKI-KmLEU2DU2500
pKI-KmLEU2DU21000
Description
Reference
E. coli and S. cerevisiae shuttle vector containing an ampicillin
resistance marker and a hisG-ScURA3-hisG blaster cassette
pKI containing a Kmku80::hisG-ScURA3-hisG disruption
cassette with a 1,000 bp homologous flanking sequence
pKI containing the Kmpdc1::hisG-ScURA3-hisG disruption
cassette with a 1,000 bp homologous flanking sequence
pKI containing the Kmpdc5::hisG-ScURA3-hisG disruption
cassette with a 1,000 bp homologous flanking sequence
pKI containing the Kmleu2:hisG-ScURA3-hisG disruption
cassette with a 50 bp homologous flanking sequence
pKI containing the KmLEU2 disruption cassette with a 100 bp
homologous flanking sequence
pKI containing the KmLEU2 disruption cassette with a 250 bp
homologous flanking sequence
pKI containing the KmLEU2 disruption cassette with a 500 bp
homologous flanking sequence
pKI containing the KmLEU2 disruption cassette with a 1,000
bp homologous flanking sequence
(Heo P et al,
2013)
6
7
8
2
this study
this study
this study
this study
this study
this study
this study
this study
9
Supplementary Table 3. List of primers used in this study
Primer name
Sequence
Description
KmKU80_1F_Xho
GCACTCGAGTTTGCACCAGGCCTTGTC
KmKU80_2B_Eco
ACTGAATTCAAACGCTGTGAGCTGGGA
KmKU80_3F_Bam
AGTGGATCCCTAATTGTATACCTGGGGC
KmKU80_4B_Sac
TGAGAGCTCGTCGAGGAATTGCTTGATC
KmPDC1_5D_1F_Xho
GATCTCGAGCCGAAAGATCGTCCGATT
KmPDC1_5D_2B_Bgl
CTAAGATCTTGCAATTATTTGGTTTGGGTG
KmPDC1_3D_3F_Xba
GCATCTAGAAGAGGGAGAGGATAAAGAG
KmPDC1_3D_4B
TCGTCTTCTCAGCTGCAA
Km05PDC6D_1F
CGTACTCGAGCCGGTGAGTCAAAGATCG
Km05PDC6D_2B
GATCGAATTCAGCTGATACCCCACCCTT
Km05PDC6D_3F
CGATACTAGTGAACTTACTCCTCAAAGC
Km05PDC6D_4B
CAGTGAGCTCATCGTGCTATCTTCCATG
Km05LEU2-5UTR_1F
CATGCTCGAGATGATGCCGTAATCAACAC
Km05LEU2-5UTR_2B
GTACAGATCTTGATGCCAAGACATTTGC
Km05LEU2-3UTR_3F
GTACACTAGTGGCTTTGACTCTAAGATTAC
Km05LEU2_3UTR_4B
TGCAGAGCTCGTGAAGCTGCTAGGCTAA
Km05Leu2-500_1FX
CATGCTCGAGAAGCACGCCCTTCTATCA
Km05Leu2+521_4BS
TGCAGAGCTCCAGAGCTTCTAAATACTCTG
Km05Leu2-261_1FX
CATGCTCGAGGCTTCAGACTCTTTGCTTG
Km05Leu2+269_4BS
TGCAGAGCTCGATATGCATACGCTTAATAA
CG
Km05Leu2-99_1FX
CATGCTCGAGGAAGTTCAAAGAATCACCA
Km05Leu2-50_1FX
CATGCTCGAGACCCACCTTTACCAATCT
ScURA3_2B_48
TTAGTTTTGCTGGCCGCA
ScURA3_1F_47
ATGTCGAAAGCTACATATAAG
Km05Leu2+103_4BS
TGCAGAGCTCCCGGAACCTAGTTTATGT
Km05Leu2+50_4BS
TGCAGAGCTCCAGCATTGAATTATTTCGTA
AT
10
3
Forward primer for amplification of
1,000 bp KmKU80 5′-UTR
Reverse primer for amplification of
1,000 bp KmKU80 5′-UTR
Forward primer for amplification of
1,000 bp KmKU80 3′-UTR
Reverse primer for amplification of
1,000 bp KmKU80 5′-UTR
Forward primer for amplification of
1,000 bp KmPDC1 5′-UTR
Reverse primer for amplification of
1,000 bp KmPDC1 5′-UTR
Forward primer for amplification of
1,000 bp KmPDC1 3′-UTR
Reverse primer for amplification of
1,000 bp KmPDC1 3′-UTR
Forward primer for amplification of
1,000 bp KmPDC5 5′-UTR
Reverse primer for amplification of
1,000 bp KmPDC5 5′UTR
Forward primer for amplification of
1,000 bp KmPDC5 3′-UTR
Reverse primer for amplification of
1,000 bp KmPDC5 3′-UTR
Forward primer for amplification of a
1,000 bp 5′-fragment of KmLEU2
Reverse primer for amplification of a
1,000 bp 5′fragment of KmLEU2
Forward primer for amplification of a
1,000 bp 3′-fragment of KmLEU2
Reverse primer for amplification of a
1,000 bp 3′-fragment of KmLEU2
Forward primer for amplification of a
500 bp 5′-fragment of KmLEU2
Reserve primer for amplification of a
500 bp 3′-fragment of KmLEU2
Forward primer for amplification of a
250 bp 5′-fragment of KmLEU2
Reserve primer for amplification of a
250 bp 3′-fragment of KmLEU2
Forward primer for amplification of a
100 bp 5′-fragment of KmLEU2
Forward primer for amplification of a 50
bp 5′-fragment of KmLEU2
Reverse primer for amplification of 100
bp and 50 bp 5′-fragments of KmLEU2
Forward primer for amplification of 100
and 50 bp 3′-fragments of KmLEU2
Reverse primer for amplification of a
100 bp 3′-fragment of KmLEU2
Reverse primer for amplification of a 50
bp 3′-fragment of KmLEU2
11
Supplementary Fig. 1 Phylogenetic tree analysis of eukaryotic Ku80 proteins including K.
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marxianus Ku80. A phylogeny was calculated using the CLUSTAL W method of the
13
DNASTAR MegAlign program. Ku80 amino acid sequences : Aspergillus niger
14
(CAK48968.1), Arabidopsis thaliana (AEE32242.1), Candida albicans (XP_713216.1),
15
Cryptococcus neoformans (XP_777203.1), Drosophila melanogaster (AAF63744.1),
16
Hansenula polymorpha (ESW97400.1), Homo sapiens (AAA36154.1), Kluyveromyces lactis
17
(AY239586.1), Penicillium chrysogenum (XP_002562026.1), Pichia pastoris (FR839631.1 ;
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region 362551-364041), Pichia stipites (XP_001383282.2), Saccharomyces cerevisiae
19
(NP_013824), Schizosaccharomyces pombe (CAC05245.1), Yarrowia lipolytica (Q6C7B9)
20
21
4
22
Supplementary Fig. 1
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24
25
5