Abstract
Two FtsZ paralogues (NbFtsZ1 and NbFtsZ2) were isolated from the unicellular green alga Nannochloris bacillaris Naumann. These sequences encoded proteins of 435 and 439 amino acids with tubulin signature motifs (GGGTG[T/S]G), which are important for GTP binding activity. NbFtsZ1 and NbFtsZ2 had four and three introns, respectively, and two different putative core promoters; a TATA box (TATAAAA) and an initiator element (CCCAGG) were located 40 bp and 80 bp upstream of the coding regions of NbFtsZ1 and NbFtsZ2, respectively. Southern blot hybridization and contourclamped homogeneous electric field electrophoresis showed that N. bacillaris contained at least one copy of each gene and that NbFtsZ1 was located on chromosome 5 and NbFtsZ2 on chromosome 3 or 4. Phylogenetically, NbFtsZ1 and NbFtsZ2 belong to the vascular plant protein families FtsZ1 and FtsZ2, respectively. The FtsZ1 proteins do not contain carboxy-terminal consensus sequences, whereas all FtsZ2 proteins possess the consensus sequence (I/V)PxFL(R/K)(K/R)(K/R). Our study has shown that NbFtsZ2 possesses a similar consensus sequence (VPDFLRRK), whereas NbFtsZ1 does not, further supporting their classification as FtsZ2 and FtsZ1. Escherichia coli ftsZ mutants transformed with cloned NbFtsZ1, and NbFtsZ2 cDNAs were restored for the capacity to divide by binary fission, suggesting that the proteins retained the ability to function in the bacterium. An anti-NbFtsZ2 anti-body specifically recognized a single protein band of approximately 51 kDa on an immunoblot of N. bacillaris cellular proteins. Immunostaining of the algal cells with this antibody produced an intense fluorescent signal as a ring near the middle of the cell, which corresponded to the chloroplast division site.
| Original language | English |
|---|---|
| Pages (from-to) | 546-556 |
| Number of pages | 11 |
| Journal | Journal of Phycology |
| Volume | 40 |
| Issue number | 3 |
| Publication status | Published - Jun 2004 |
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Keywords
- Chloroplast division
- FtsZ
- Nannochloris bacillaris
ASJC Scopus subject areas
- Aquatic Science
- Plant Science
Cite this
Molecular divergence and characterization of two chloroplast division genes, FtsZ1 and FtsZ2, in the unicellular green alga Nannochloris bacillaris (Chlorophyta). / Koide, Tomojirou; Yamazaki, Tomokazu; Yamamoto, Maki; Fujishita, Mariko; Nomura, Hideo; Moriyama, Yohsuke; Sumiya, Nobuko; Matsunaga, Sachihiro; Sakamoto, Wataru; Kawano, Shigeyuki.
In: Journal of Phycology, Vol. 40, No. 3, 06.2004, p. 546-556.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Molecular divergence and characterization of two chloroplast division genes, FtsZ1 and FtsZ2, in the unicellular green alga Nannochloris bacillaris (Chlorophyta)
AU - Koide, Tomojirou
AU - Yamazaki, Tomokazu
AU - Yamamoto, Maki
AU - Fujishita, Mariko
AU - Nomura, Hideo
AU - Moriyama, Yohsuke
AU - Sumiya, Nobuko
AU - Matsunaga, Sachihiro
AU - Sakamoto, Wataru
AU - Kawano, Shigeyuki
PY - 2004/6
Y1 - 2004/6
N2 - Two FtsZ paralogues (NbFtsZ1 and NbFtsZ2) were isolated from the unicellular green alga Nannochloris bacillaris Naumann. These sequences encoded proteins of 435 and 439 amino acids with tubulin signature motifs (GGGTG[T/S]G), which are important for GTP binding activity. NbFtsZ1 and NbFtsZ2 had four and three introns, respectively, and two different putative core promoters; a TATA box (TATAAAA) and an initiator element (CCCAGG) were located 40 bp and 80 bp upstream of the coding regions of NbFtsZ1 and NbFtsZ2, respectively. Southern blot hybridization and contourclamped homogeneous electric field electrophoresis showed that N. bacillaris contained at least one copy of each gene and that NbFtsZ1 was located on chromosome 5 and NbFtsZ2 on chromosome 3 or 4. Phylogenetically, NbFtsZ1 and NbFtsZ2 belong to the vascular plant protein families FtsZ1 and FtsZ2, respectively. The FtsZ1 proteins do not contain carboxy-terminal consensus sequences, whereas all FtsZ2 proteins possess the consensus sequence (I/V)PxFL(R/K)(K/R)(K/R). Our study has shown that NbFtsZ2 possesses a similar consensus sequence (VPDFLRRK), whereas NbFtsZ1 does not, further supporting their classification as FtsZ2 and FtsZ1. Escherichia coli ftsZ mutants transformed with cloned NbFtsZ1, and NbFtsZ2 cDNAs were restored for the capacity to divide by binary fission, suggesting that the proteins retained the ability to function in the bacterium. An anti-NbFtsZ2 anti-body specifically recognized a single protein band of approximately 51 kDa on an immunoblot of N. bacillaris cellular proteins. Immunostaining of the algal cells with this antibody produced an intense fluorescent signal as a ring near the middle of the cell, which corresponded to the chloroplast division site.
AB - Two FtsZ paralogues (NbFtsZ1 and NbFtsZ2) were isolated from the unicellular green alga Nannochloris bacillaris Naumann. These sequences encoded proteins of 435 and 439 amino acids with tubulin signature motifs (GGGTG[T/S]G), which are important for GTP binding activity. NbFtsZ1 and NbFtsZ2 had four and three introns, respectively, and two different putative core promoters; a TATA box (TATAAAA) and an initiator element (CCCAGG) were located 40 bp and 80 bp upstream of the coding regions of NbFtsZ1 and NbFtsZ2, respectively. Southern blot hybridization and contourclamped homogeneous electric field electrophoresis showed that N. bacillaris contained at least one copy of each gene and that NbFtsZ1 was located on chromosome 5 and NbFtsZ2 on chromosome 3 or 4. Phylogenetically, NbFtsZ1 and NbFtsZ2 belong to the vascular plant protein families FtsZ1 and FtsZ2, respectively. The FtsZ1 proteins do not contain carboxy-terminal consensus sequences, whereas all FtsZ2 proteins possess the consensus sequence (I/V)PxFL(R/K)(K/R)(K/R). Our study has shown that NbFtsZ2 possesses a similar consensus sequence (VPDFLRRK), whereas NbFtsZ1 does not, further supporting their classification as FtsZ2 and FtsZ1. Escherichia coli ftsZ mutants transformed with cloned NbFtsZ1, and NbFtsZ2 cDNAs were restored for the capacity to divide by binary fission, suggesting that the proteins retained the ability to function in the bacterium. An anti-NbFtsZ2 anti-body specifically recognized a single protein band of approximately 51 kDa on an immunoblot of N. bacillaris cellular proteins. Immunostaining of the algal cells with this antibody produced an intense fluorescent signal as a ring near the middle of the cell, which corresponded to the chloroplast division site.
KW - Chloroplast division
KW - FtsZ
KW - Nannochloris bacillaris
UR - http://www.scopus.com/inward/record.url?scp=2942628006&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=2942628006&partnerID=8YFLogxK
M3 - Article
AN - SCOPUS:2942628006
VL - 40
SP - 546
EP - 556
JO - Journal of Phycology
JF - Journal of Phycology
SN - 0022-3646
IS - 3
ER -