Tandem affinity purification of AtTERT reveals putative
interaction partners of plant telomerase in vivo

J 2017

Tandem affinity purification of AtTERT reveals putative interaction partners of plant telomerase in vivo

MAJERSKÁ, Jana, Petra PROCHÁZKOVÁ SCHRUMPFOVÁ, Ladislav DOKLADAL, Šárka SCHOŘOVÁ, Karel STEJSKAL et. al.

Basic information

Original name

Tandem affinity purification of AtTERT reveals putative interaction partners of plant telomerase in vivo

Authors

MAJERSKÁ, Jana (703 Slovakia, belonging to the institution), Petra PROCHÁZKOVÁ SCHRUMPFOVÁ (203 Czech Republic, belonging to the institution), Ladislav DOKLADAL (203 Czech Republic), Šárka SCHOŘOVÁ (203 Czech Republic, guarantor, belonging to the institution), Karel STEJSKAL (203 Czech Republic, belonging to the institution), Michal OBOŘIL (203 Czech Republic, belonging to the institution), David HONYS (203 Czech Republic), Lucie KOZÁKOVÁ (203 Czech Republic, belonging to the institution), Pavla SOVÁKOVÁ (203 Czech Republic, belonging to the institution) and Eva SYKOROVA (203 Czech Republic)

Edition

Protoplasma, Wien, Springer-Verlag, 2017, 0033-183X

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10600 1.6 Biological sciences

Country of publisher

Austria

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 2.457

RIV identification code

RIV/00216224:14740/17:00094874

Organization unit

Central European Institute of Technology

DOI

http://dx.doi.org/10.1007/s00709-016-1042-3

UT WoS

000403774000009

Keywords in English

Telomerase; TAP-MS; AtPOT1a; Pontin; Reptin; PUR alpha 1

Abstract

V originále

The life cycle of telomerase involves dynamic and complex interactions between proteins within multiple macromolecular networks. Elucidation of these associations is a key to understanding the regulation of telomerase under diverse physiological and pathological conditions from telomerase biogenesis, through telomere recruitment and elongation, to its non-canonical activities outside of telomeres. We used tandem affinity purification coupled to mass spectrometry to build an interactome of the telomerase catalytic subunit AtTERT, using Arabidopsis thaliana suspension cultures. We then examined interactions occurring at the AtTERT N-terminus, which is thought to fold into a discrete domain connected to the rest of the molecule via a flexible linker. Bioinformatic analyses revealed that interaction partners of AtTERT have a range of molecular functions, a subset of which is specific to the network around its N-terminus. A significant number of proteins co-purifying with the N-terminal constructs have been implicated in cell cycle and developmental processes, as would be expected of bona fide regulatory interactions and we have confirmed experimentally the direct nature of selected interactions. To examine AtTERT protein-protein interactions from another perspective, we also analysed AtTERT interdomain contacts to test potential dimerization of AtTERT. In total, our results provide an insight into the composition and architecture of the plant telomerase complex and this will aid in delineating molecular mechanisms of telomerase functions.

Links

GA13-06943S, research and development project

Name: Strukturní a funkční komponenty rostlinných telomer

Investor: Czech Science Foundation

LQ1601, research and development project

Name: CEITEC 2020 (Acronym: CEITEC2020)

Investor: Ministry of Education, Youth and Sports of the CR

MUNI/C/0981/2010, interní kód MU

Name: Funkční charakterizace domén rostlinné telomerázy (Acronym: TAPTERT)

Investor: Masaryk University, Rector's Program

Citovat

MAJERSKÁ, Jana, Petra PROCHÁZKOVÁ SCHRUMPFOVÁ, Ladislav DOKLADAL, Šárka SCHOŘOVÁ, Karel STEJSKAL, Michal OBOŘIL, David HONYS, Lucie KOZÁKOVÁ, Pavla SOVÁKOVÁ and Eva SYKOROVA. Tandem affinity purification of AtTERT reveals putative interaction partners of plant telomerase in vivo. Protoplasma. Wien: Springer-Verlag, 2017, vol. 254, No 4, p. 1547-1562. ISSN 0033-183X. Available from: https://dx.doi.org/10.1007/s00709-016-1042-3.

@article{1386747,
   author = {Majerská, Jana and Procházková Schrumpfová, Petra and Dokladal, Ladislav and Schořová, Šárka and Stejskal, Karel and Obořil, Michal and Honys, David and Kozáková, Lucie and Sováková, Pavla and Sykorova, Eva},
   article_location = {Wien},
   article_number = {4},
   doi = {http://dx.doi.org/10.1007/s00709-016-1042-3},
   keywords = {Telomerase; TAP-MS; AtPOT1a; Pontin; Reptin; PUR alpha 1},
   language = {eng},
   issn = {0033-183X},
   journal = {Protoplasma},
   title = {Tandem affinity purification of AtTERT reveals putative interaction partners of plant telomerase in vivo},
   url = {https://link.springer.com/article/10.1007%2Fs00709-016-1042-3},
   volume = {254},
   year = {2017}
}

TY  - JOUR
ID  - 1386747
AU  - Majerská, Jana - Procházková Schrumpfová, Petra - Dokladal, Ladislav - Schořová, Šárka - Stejskal, Karel - Obořil, Michal - Honys, David - Kozáková, Lucie - Sováková, Pavla - Sykorova, Eva
PY  - 2017
TI  - Tandem affinity purification of AtTERT reveals putative interaction partners of plant telomerase in vivo
JF  - Protoplasma
VL  - 254
IS  - 4
SP  - 1547-1562
EP  - 1547-1562
PB  - Springer-Verlag
SN  - 0033183X
KW  - Telomerase
KW  - TAP-MS
KW  - AtPOT1a
KW  - Pontin
KW  - Reptin
KW  - PUR alpha 1
UR  - https://link.springer.com/article/10.1007%2Fs00709-016-1042-3
L2  - https://link.springer.com/article/10.1007%2Fs00709-016-1042-3
N2  - The life cycle of telomerase involves dynamic and complex interactions between proteins within multiple macromolecular networks. Elucidation of these associations is a key to understanding the regulation of telomerase under diverse physiological and pathological conditions from telomerase biogenesis, through telomere recruitment and elongation, to its non-canonical activities outside of telomeres. We used tandem affinity purification coupled to mass spectrometry to build an interactome of the telomerase catalytic subunit AtTERT, using Arabidopsis thaliana suspension cultures. We then examined interactions occurring at the AtTERT N-terminus, which is thought to fold into a discrete domain connected to the rest of the molecule via a flexible linker. Bioinformatic analyses revealed that interaction partners of AtTERT have a range of molecular functions, a subset of which is specific to the network around its N-terminus. A significant number of proteins co-purifying with the N-terminal constructs have been implicated in cell cycle and developmental processes, as would be expected of bona fide regulatory interactions and we have confirmed experimentally the direct nature of selected interactions. To examine AtTERT protein-protein interactions from another perspective, we also analysed AtTERT interdomain contacts to test potential dimerization of AtTERT. In total, our results provide an insight into the composition and architecture of the plant telomerase complex and this will aid in delineating molecular mechanisms of telomerase functions.
ER  -

MAJERSKÁ, Jana, Petra PROCHÁZKOVÁ SCHRUMPFOVÁ, Ladislav DOKLADAL, Šárka SCHOŘOVÁ, Karel STEJSKAL, Michal OBOŘIL, David HONYS, Lucie KOZÁKOVÁ, Pavla SOVÁKOVÁ and Eva SYKOROVA. Tandem affinity purification of AtTERT reveals putative interaction partners of plant telomerase in vivo. \textit{Protoplasma}. Wien: Springer-Verlag, 2017, vol.~254, No~4, p.~1547-1562. ISSN~0033-183X. Available from: https://dx.doi.org/10.1007/s00709-016-1042-3.

Detailed Information on Publication Record