PMID- 30530494
OWN - NLM
STAT- MEDLINE
DCOM- 20190520
LR  - 20210314
IS  - 1083-351X (Electronic)
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 294
IP  - 5
DP  - 2019 Feb 1
TI  - An interdomain bridge influences RNA binding of the human La protein.
PG  - 1529-1540
LID - S0021-9258(20)36463-2 [pii]
LID - 10.1074/jbc.RA118.003995 [doi]
AB  - La proteins are RNA chaperones that perform various functions depending on 
      distinct RNA-binding modes and their subcellular localization. In the nucleus, 
      they help process UUU-3'OH-tailed nascent RNA polymerase III transcripts, such as 
      pre-tRNAs, whereas in the cytoplasm they contribute to translation of 
      poly(A)-tailed mRNAs. La accumulation in the nucleus and cytoplasm is controlled 
      by several trafficking elements, including a canonical nuclear localization 
      signal in the extreme C terminus and a nuclear retention element (NRE) in the RNA 
      recognition motif 2 (RRM2) domain. Previous findings indicate that cytoplasmic 
      export of La due to mutation of the NRE can be suppressed by mutations in RRM1, 
      but the mechanism by which the RRM1 and RRM2 domains functionally cooperate is 
      poorly understood. In this work, we use electromobility shift assays (EMSA) to 
      show that mutations in the NRE and RRM1 affect binding of human La to pre-tRNAs 
      but not UUU-3'OH or poly(A) sequences, and we present compensatory mutagenesis 
      data supporting a direct interaction between the RRM1 and RRM2 domains. Moreover, 
      we use collision-induced unfolding and time-resolved hydrogen-deuterium exchange 
      MS analyses to study the conformational dynamics that occur when this interaction 
      is intact or disrupted. Our results suggest that the intracellular distribution 
      of La may be linked to its RNA-binding modes and provide the first evidence for a 
      direct protein-protein interdomain interaction in La proteins.
CI  - (c) 2019 Marrella et al.
FAU - Marrella, Stefano A
AU  - Marrella SA
AD  - Department of Biology, York University, Toronto, Ontario M3J 1P3, Canada; Centres 
      for Research in Biomolecular Interactions, York University, Toronto, Ontario M3J 
      1P3, Canada.
FAU - Brown, Kerene A
AU  - Brown KA
AD  - Centres for Research in Biomolecular Interactions, York University, Toronto, 
      Ontario M3J 1P3, Canada; Department of Chemistry, York University, Toronto, 
      Ontario M3J 1P3, Canada; Research in Mass Spectrometry, York University, Toronto, 
      Ontario M3J 1P3, Canada.
FAU - Mansouri-Noori, Farnaz
AU  - Mansouri-Noori F
AD  - Department of Biology, York University, Toronto, Ontario M3J 1P3, Canada; Centres 
      for Research in Biomolecular Interactions, York University, Toronto, Ontario M3J 
      1P3, Canada.
FAU - Porat, Jennifer
AU  - Porat J
AD  - Department of Biology, York University, Toronto, Ontario M3J 1P3, Canada; Centres 
      for Research in Biomolecular Interactions, York University, Toronto, Ontario M3J 
      1P3, Canada.
FAU - Wilson, Derek J
AU  - Wilson DJ
AD  - Centres for Research in Biomolecular Interactions, York University, Toronto, 
      Ontario M3J 1P3, Canada; Department of Chemistry, York University, Toronto, 
      Ontario M3J 1P3, Canada; Research in Mass Spectrometry, York University, Toronto, 
      Ontario M3J 1P3, Canada. Electronic address: dkwilson@yorku.ca.
FAU - Bayfield, Mark A
AU  - Bayfield MA
AD  - Department of Biology, York University, Toronto, Ontario M3J 1P3, Canada; Centres 
      for Research in Biomolecular Interactions, York University, Toronto, Ontario M3J 
      1P3, Canada. Electronic address: bayfield@yorku.ca.
LA  - eng
SI  - PDB/2VOO
SI  - PDB/1OWX
SI  - PDB/1YTY
GR  - CIHR/Canada
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20181210
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (La protein, human)
RN  - 0 (Phosphoproteins)
RN  - 63231-63-0 (RNA)
SB  - IM
MH  - Binding Sites
MH  - Cell Nucleus/genetics/*metabolism
MH  - Humans
MH  - Models, Molecular
MH  - Mutation
MH  - Phosphoproteins/*chemistry/genetics/*metabolism
MH  - Protein Binding
MH  - Protein Conformation
MH  - Protein Domains
MH  - Protein Folding
MH  - RNA/chemistry/*metabolism
MH  - *RNA Recognition Motif
PMC - PMC6364776
OTO - NOTNLM
OT  - La protein
OT  - RNA processing
OT  - RNA-binding protein
OT  - RNA-protein interaction
OT  - SSB
OT  - Sjogren syndrome antigen B
OT  - intracellular trafficking
OT  - mass spectrometry (MS)
OT  - precursor tRNA (pre-tRNA)
OT  - protein domain
COIS- The authors declare that they have no conflicts of interest with the contents of 
      this article
EDAT- 2018/12/12 06:00
MHDA- 2019/05/21 06:00
PMCR- 2020/02/01
CRDT- 2018/12/12 06:00
PHST- 2018/05/23 00:00 [received]
PHST- 2018/11/20 00:00 [revised]
PHST- 2018/12/12 06:00 [pubmed]
PHST- 2019/05/21 06:00 [medline]
PHST- 2018/12/12 06:00 [entrez]
PHST- 2020/02/01 00:00 [pmc-release]
AID - S0021-9258(20)36463-2 [pii]
AID - RA118.003995 [pii]
AID - 10.1074/jbc.RA118.003995 [doi]
PST - ppublish
SO  - J Biol Chem. 2019 Feb 1;294(5):1529-1540. doi: 10.1074/jbc.RA118.003995. Epub 
      2018 Dec 10.