PMID- 24854174
OWN - NLM
STAT- MEDLINE
DCOM- 20140921
LR  - 20191210
IS  - 1470-8728 (Electronic)
IS  - 0264-6021 (Linking)
VI  - 461
IP  - 3
DP  - 2014 Aug 1
TI  - Overlapping binding sites for importin beta1 and suppressor of fused (SuFu) on 
      glioma-associated oncogene homologue 1 (Gli1) regulate its nuclear localization.
PG  - 469-76
LID - 10.1042/BJ20130709 [doi]
AB  - A key factor in oncogenesis is the transport into the nucleus of oncogenic 
      signalling molecules, such as Gli1 (glioma-associated oncogene homologue 1), the 
      central transcriptional activator in the Hedgehog signalling pathway. Little is 
      known, however, how factors such as Gli are transported into the nucleus and how 
      this may be regulated by interaction with other cellular factors, such as the 
      negative regulator suppressor of fused (SuFu). In the present study we show for 
      the first time that nuclear entry of Gli1 is regulated by a unique mechanism 
      through mutually exclusive binding by its nuclear import factor Impbeta1 (importin 
      beta1) and SuFu. Using quantitative live mammalian cell imaging, we show that 
      nuclear accumulation of GFP-Gli1 fusion proteins, but not of a control protein, 
      is specifically inhibited by co-expression of SuFu. Using a direct binding assay, 
      we show that Impbeta1 exhibits a high nanomolar affinity to Gli1, with specific 
      knockdown of Impbeta1 expression being able to inhibit Gli1 nuclear accumulation, 
      thus implicating Impbeta1 as the nuclear transporter for Gli1 for the first time. 
      SuFu also binds to Gli1 with a high nanomolar affinity, intriguingly being able 
      to compete with Impbeta1 for binding to Gli1, through the fact that the sites for 
      SuFu and Impbeta1 binding overlap at the Gli1 N-terminus. The results indicate for 
      the first time that the relative intracellular concentrations of SuFu and Impbeta1 
      are likely to determine the localization of Gli1, with implications for its 
      action in cancer, as well as in developmental systems.
FAU - Szczepny, Anette
AU  - Szczepny A
FAU - Wagstaff, Kylie M
AU  - Wagstaff KM
AD  -  section signDepartment of Biochemistry and Molecular Biology, Monash University, Melbourne, 
      Victoria 3800, Australia.
FAU - Dias, Manisha
AU  - Dias M
AD  -  section signDepartment of Biochemistry and Molecular Biology, Monash University, Melbourne, 
      Victoria 3800, Australia.
FAU - Gajewska, Katarzyna
AU  - Gajewska K
AD  -  section signDepartment of Biochemistry and Molecular Biology, Monash University, Melbourne, 
      Victoria 3800, Australia.
FAU - Wang, Chunxiao
AU  - Wang C
AD  -  section signDepartment of Biochemistry and Molecular Biology, Monash University, Melbourne, 
      Victoria 3800, Australia.
FAU - Davies, Rebecca G
AU  - Davies RG
AD  -  section signDepartment of Biochemistry and Molecular Biology, Monash University, Melbourne, 
      Victoria 3800, Australia.
FAU - Kaur, Gurpreet
AU  - Kaur G
AD  -  section signDepartment of Biochemistry and Molecular Biology, Monash University, Melbourne, 
      Victoria 3800, Australia.
FAU - Ly-Huynh, Jennifer
AU  - Ly-Huynh J
FAU - Loveland, Kate L
AU  - Loveland KL
FAU - Jans, David A
AU  - Jans DA
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Biochem J
JT  - The Biochemical journal
JID - 2984726R
RN  - 0 (GLI1 protein, human)
RN  - 0 (Luminescent Proteins)
RN  - 0 (Neoplasm Proteins)
RN  - 0 (Peptide Fragments)
RN  - 0 (RNA, Small Interfering)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (Repressor Proteins)
RN  - 0 (Sufu protein, mouse)
RN  - 0 (Transcription Factors)
RN  - 0 (Zinc Finger Protein GLI1)
RN  - 0 (beta Karyopherins)
SB  - IM
MH  - Adenocarcinoma/*metabolism/pathology
MH  - Animals
MH  - Binding Sites
MH  - Binding, Competitive
MH  - COS Cells
MH  - Cell Nucleus/*metabolism/ultrastructure
MH  - Chlorocebus aethiops
MH  - HeLa Cells
MH  - Humans
MH  - Luminescent Proteins/genetics/metabolism
MH  - Neoplasm Proteins/antagonists & inhibitors/chemistry/genetics/*metabolism
MH  - Peptide Fragments/chemistry/genetics/metabolism
MH  - Protein Interaction Domains and Motifs
MH  - Protein Transport
MH  - RNA Interference
MH  - RNA, Small Interfering
MH  - Recombinant Fusion Proteins/chemistry/metabolism
MH  - Repressor Proteins/chemistry/genetics/*metabolism
MH  - Transcription Factors/chemistry/genetics/*metabolism
MH  - Zinc Finger Protein GLI1
MH  - beta Karyopherins/antagonists & inhibitors/chemistry/genetics/*metabolism
EDAT- 2014/05/24 06:00
MHDA- 2014/09/23 06:00
CRDT- 2014/05/24 06:00
PHST- 2014/05/24 06:00 [entrez]
PHST- 2014/05/24 06:00 [pubmed]
PHST- 2014/09/23 06:00 [medline]
AID - BJ20130709 [pii]
AID - 10.1042/BJ20130709 [doi]
PST - ppublish
SO  - Biochem J. 2014 Aug 1;461(3):469-76. doi: 10.1042/BJ20130709.