GARBARINO PICO EDUARDO
Congresos y reuniones científicas
Título:
The circadian deadenylase Nocturnin interacts with a novel form of the RNA-binding protein KSRP
Autor/es:
GARBARINO PICO E, NIU S AND GREEN CB
Lugar:
Tucumán, Argentina
Reunión:
Congreso; XLV Reunión Anual Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2009
Institución organizadora:
SAIB
Resumen:
CB-P32.
THE CIRCADIAN DEADENYLASE NOCTURNIN
INTERACTS WITH A NOVEL FORM OF THE RNABINDING
PROTEIN KSRP
Garbarino Pico E1; Niu S2; Green CB2
1CIQUIBIC-Dpto. Química Biológica, Facultad Ciencias
Químicas, UNCórdoba. 2University of Virginia. E-mail:
garba@mail.fcq.unc.edu.ar
Mouse nocturnin (mNOC) is a circadian regulated deadenylase.
mRNA deadenylation induces transcript degradation or
translational silencing. In order to identify mNOC binding partners,
we performed a large-scale immunoprecipitation (IP) screen. The
mass spectroscopy analysis of co-IP proteins revealed KSRP (KH
homology splicing regulatory protein) as a potential mNOC binding
protein. KSRP is an AU-rich element (ARE) containing mRNA
binding protein that regulates mRNA turnover. We verified this
interaction by co-IP. We utilized three anti-KSRP antibodies, two of
them detected two KSRP forms with apparent molecular weigh of
75 and 60 kDa, whereas the other one (monoclonal), only the 75 kDa
form. mNOC co-IP and co-fractionate in gel filtration with the 60
kDa form. Since KSRP is also involved in nuclear splicing, we
studied the mNOC and KSRP60 subcellular localization to
determine where this interaction takes place. ICC and biochemical
fractionation showed that both proteins co-localize in cytoplasm,
whereas KSRP75 is in the nucleus. We confirmed the identity of
KSRP60 by repeating the mass spectroscopy analysis.
Our results suggest that NOC may play a role in regulating ARE-mRNA
metabolism. Our current working hypothesis is that mNOC
is recruited to ARE-mRNAs by KSRP60, which results in the
deadenylation of these messages, and consequently, their
degradation or translational silencing.
THE CIRCADIAN DEADENYLASE NOCTURNIN
INTERACTS WITH A NOVEL FORM OF THE RNABINDING
PROTEIN KSRP
Garbarino Pico E1; Niu S2; Green CB2
1CIQUIBIC-Dpto. Química Biológica, Facultad Ciencias
Químicas, UNCórdoba. 2University of Virginia. E-mail:
garba@mail.fcq.unc.edu.ar
Mouse nocturnin (mNOC) is a circadian regulated deadenylase.
mRNA deadenylation induces transcript degradation or
translational silencing. In order to identify mNOC binding partners,
we performed a large-scale immunoprecipitation (IP) screen. The
mass spectroscopy analysis of co-IP proteins revealed KSRP (KH
homology splicing regulatory protein) as a potential mNOC binding
protein. KSRP is an AU-rich element (ARE) containing mRNA
binding protein that regulates mRNA turnover. We verified this
interaction by co-IP. We utilized three anti-KSRP antibodies, two of
them detected two KSRP forms with apparent molecular weigh of
75 and 60 kDa, whereas the other one (monoclonal), only the 75 kDa
form. mNOC co-IP and co-fractionate in gel filtration with the 60
kDa form. Since KSRP is also involved in nuclear splicing, we
studied the mNOC and KSRP60 subcellular localization to
determine where this interaction takes place. ICC and biochemical
fractionation showed that both proteins co-localize in cytoplasm,
whereas KSRP75 is in the nucleus. We confirmed the identity of
KSRP60 by repeating the mass spectroscopy analysis.
Our results suggest that NOC may play a role in regulating ARE-mRNA
metabolism. Our current working hypothesis is that mNOC
is recruited to ARE-mRNAs by KSRP60, which results in the
deadenylation of these messages, and consequently, their
degradation or translational silencing.
