Export Receptors | ScienceGate

ABSTRACT Ribosomal maturation is a complex and highly conserved biological process involving migration of a continuously changing RNP across multiple cellular compartments. A critical point in this process is the translocation of individual ribosomal subunits (60S and 40S) from the nucleus to the cytoplasm, and a number of export factors participate in this process. In this study, we characterize the functional role of the auxiliary export receptors TbMex67 and TbMtr2 in ribosome biogenesis in the parasite Trypanosoma brucei. We demonstrate that depletion of each of these proteins dramatically impacts the steady-state levels of other proteins involved in ribosome biogenesis, including the trypanosome-specific factors P34 and P37. In addition, we observe that the loss of TbMex67 or TbMtr2 leads to aberrant ribosome formation, rRNA processing, and polysome formation. Although the TbMex67-TbMtr2 heterodimer is structurally distinct from Mex67-Mtr2 complexes previously studied, our data show that they retain a conserved function in ribosome biogenesis. IMPORTANCE The nuclear export of ribosomal subunits (60S and 40S) depends in part on the activity of the essential auxiliary export receptors TbMtr2 and TbMex67. When these proteins are individually depleted from the medically and agriculturally significant parasite Trypanosoma brucei, distinct alterations in the processing of the rRNAs of the large subunit (60S) are observed as well as aberrations in the assembly of functional ribosomes (polysomes). We also established that TbMex67 and TbMtr2 interact directly or indirectly with the protein components of the 5S RNP, including the trypanosome-specific P34/P37. The critical role that TbMex67 and TbMtr2 play in this essential biological process together with their parasite-specific interactions may provide new therapeutic targets against this important parasite.

Download Full-text

"Friedrich Miescher Prize Awardee Lecture Review"¶A conserved family of nuclear export receptors mediates the exit of messenger RNA to the cytoplasm

Cellular and Molecular Life Sciences ◽

10.1007/pl00000924 ◽

2001 ◽

Vol 58 (8) ◽

pp. 1105-1112 ◽

Cited By ~ 10

Author(s):

E. Izaurralde

Keyword(s):

Nuclear Export ◽

Messenger Rna ◽

Export Receptors

Download Full-text

Utp22p acts in concert with Utp8p to channel aminoacyl-tRNA from the nucleolus to the nuclear tRNA export receptor Los1p but not Msn5p

Biochemistry and Cell Biology ◽

10.1139/o2012-034 ◽

2012 ◽

Vol 90 (6) ◽

pp. 731-749 ◽

Cited By ~ 4

Author(s):

Manoja B.K. Eswara ◽

Ashley Clayton ◽

Dev Mangroo

Keyword(s):

Protein Complex ◽

Trna Synthetase ◽

Nuclear Pore ◽

Independent Manner ◽

Nuclear Retention ◽

Trna Synthetases ◽

U3 Snorna ◽

Aminoacyl Trna Synthetases ◽

Export Receptors

Utp8p is an essential nucleolar protein that channels aminoacyl-tRNAs from aminoacyl-tRNA synthetases in the nucleolus to the nuclear tRNA export receptors located in the nucleoplasm and nuclear pore complex in Saccharomyces cerevisiae . Utp8p is also part of the U3 snoRNA-associated protein complex involved in 18S rRNA biogenesis in the nucleolus. We report that Utp22p, which is another member of the U3 snoRNA-associated protein complex, is also an intranuclear component of the nuclear tRNA export machinery. Depletion of Utp22p results in nuclear retention of mature tRNAs derived from intron-containing and intronless precursors. Moreover, Utp22p copurifies with the nuclear tRNA export receptor Los1p, the aminoacyl-tRNA synthetase Tys1p and Utp8p, but not with the RanGTPase Gsp1p and the nuclear tRNA export receptor Msn5p. Utp22p interacts directly with Utp8p and Los1p in a tRNA-independent manner in vitro. Utp22p also interacts directly with Tys1p, but this binding is stimulated when Tys1p is bound to tRNA. However, Utp22p, unlike Utp8p, does not bind tRNA saturably. These data suggest that Utp22p recruits Utp8p to aminoacyl-tRNA synthetases in the nucleolus to collect aminoacyl-tRNA and then accompanies the Utp8p–tRNA complex to deliver the aminoacyl-tRNAs to Los1p but not Msn5p. It is possible that Nrap/Nol6, the mammalian orthologue of Utp22p, plays a role in channelling aminoacyl-tRNA to the nuclear tRNA export receptor exportin-t.

Download Full-text

Utp9p Facilitates Msn5p-mediated Nuclear Reexport of Retrograded tRNAs in Saccharomyces cerevisiae

Molecular Biology of the Cell ◽

10.1091/mbc.e09-06-0490 ◽

2009 ◽

Vol 20 (23) ◽

pp. 5007-5025 ◽

Cited By ~ 23

Author(s):

Manoja B.K. Eswara ◽

Andrew T. McGuire ◽

Jacqueline B. Pierce ◽

Dev Mangroo

Keyword(s):

Saccharomyces Cerevisiae ◽

Nuclear Accumulation ◽

Dependent Manner ◽

Trna Synthetases ◽

Trna Splicing ◽

U3 Snorna ◽

Aminoacyl Trna Synthetases ◽

Associated Proteins ◽

Export Receptors

Utp9p is a nucleolar protein that is part of a subcomplex containing several U3 snoRNA-associated proteins including Utp8p, which is a protein that shuttles aminoacyl-tRNAs from the nucleolus to the nuclear tRNA export receptors Los1p and Msn5p in Saccharomyces cerevisiae . Here we show that Utp9p is also an intranuclear component of the Msn5p-mediated nuclear tRNA export pathway. Depletion of Utp9p caused nuclear accumulation of mature tRNAs derived from intron-containing precursors, but not tRNAs made from intronless pre-tRNAs. Utp9p binds tRNA directly and saturably, and copurifies with Utp8p, Gsp1p, and Msn5p, but not with Los1p or aminoacyl-tRNA synthetases. Utp9p interacts directly with Utp8p, Gsp1p, and Msn5p in vitro. Furthermore, Gsp1p forms a complex with Msn5p and Utp9p in a tRNA-dependent manner. However, Utp9p does not shuttle between the nucleus and the cytoplasm. Because tRNA splicing occurs in the cytoplasm and the spliced tRNAs are retrograded back to the nucleus, we propose that Utp9p facilitates nuclear reexport of retrograded tRNAs. Moreover, the data suggest that Utp9p together with Utp8p translocate aminoacyl-tRNAs from the nucleolus to Msn5p and assist with formation of the Msn5p-tRNA-Gsp1p-GTP export complex.

Download Full-text

Protein Kinase A Is Part of a Mechanism That Regulates Nuclear Reimport of the Nuclear tRNA Export Receptors Los1p and Msn5p

Eukaryotic Cell ◽

10.1128/ec.00214-13 ◽

2013 ◽

Vol 13 (2) ◽

pp. 209-230 ◽

Cited By ~ 6

Author(s):

Jacqueline B. Pierce ◽

George van der Merwe ◽

Dev Mangroo

Keyword(s):

Signal Transduction ◽

Protein Kinase ◽

Protein Kinase A ◽

Signal Transduction Pathway ◽

Glucose Deprivation ◽

Content Type ◽

Cytoplasmic Accumulation ◽

Export Receptors ◽

Kinase A ◽

Glucose Availability

ABSTRACTThe two main signal transduction mechanisms that allow eukaryotes to sense and respond to changes in glucose availability in the environment are the cyclic AMP (cAMP)/protein kinase A (PKA) and AMP-activated protein kinase (AMPK)/Snf1 kinase-dependent pathways. Previous studies have shown that the nuclear tRNA export process is inhibited inSaccharomyces cerevisiaedeprived of glucose. However, the signal transduction pathway involved and the mechanism by which glucose availability regulates nuclear-cytoplasmic tRNA trafficking are not understood. Here, we show that inhibition of nuclear tRNA export is caused by a block in nuclear reimport of the tRNA export receptors during glucose deprivation. Cytoplasmic accumulation of the tRNA export receptors during glucose deprivation is not caused by activation of Snf1p. Evidence obtained suggests that PKA is part of the mechanism that regulates nuclear reimport of the tRNA export receptors in response to glucose availability. This mechanism does not appear to involve phosphorylation of the nuclear tRNA export receptors by PKA. The block in nuclear reimport of the tRNA export receptors appears to be caused by activation of an unidentified mechanism when PKA is turned off during glucose deprivation. Taken together, the data suggest that PKA facilitates return of the tRNA export receptors to the nucleus by inhibiting an unidentified activity that facilitates cytoplasmic accumulation of the tRNA export receptors when glucose in the environment is limiting. A PKA-independent mechanism was also found to regulate nuclear tRNA export in response to glucose availability. This mechanism, however, does not regulate nuclear reimport of the tRNA export receptors.

Download Full-text

Regulation of RNA-binding proteins affinity to export receptors enables the nuclear basket proteins to distinguish and retain aberrant mRNAs

Scientific Reports ◽

10.1038/srep35380 ◽

2016 ◽

Vol 6 (1) ◽

Cited By ~ 5

Author(s):

M. Soheilypour ◽

M. R. K. Mofrad

Keyword(s):

Binding Proteins ◽

Rna Binding ◽

Rna Binding Proteins ◽

Export Receptors

Download Full-text

Observation of Single Nuclear Export Receptors at the Nuclear Pore Complex

Biophysical Journal ◽

10.1016/j.bpj.2009.12.1670 ◽

2010 ◽

Vol 98 (3) ◽

pp. 307a

Author(s):

Ulrike Schmitz-Ziffels ◽

Andreas Veenendaal ◽

Jan Peter Siebrasse ◽

Ulrich Kubitscheck

Keyword(s):

Nuclear Pore Complex ◽

Nuclear Export ◽

Nuclear Pore ◽

Export Receptors

Download Full-text

Arx1 Is a Nuclear Export Receptor for the 60S Ribosomal Subunit in Yeast

Molecular Biology of the Cell ◽

10.1091/mbc.e07-09-0968 ◽

2008 ◽

Vol 19 (2) ◽

pp. 735-744 ◽

Cited By ~ 57

Author(s):

Nai-Jung Hung ◽

Kai-Yin Lo ◽

Samir S. Patel ◽

Kara Helmke ◽

Arlen W. Johnson

Keyword(s):

Nuclear Export ◽

Ribosomal Subunit ◽

In Vitro Assays ◽

Dependent Manner ◽

Yeast Two Hybrid ◽

Synthetic Lethal ◽

Nuclear Export Receptor ◽

Export Receptors ◽

Two Hybrid

We previously showed that nuclear export of the large (60S) ribosomal subunit relies on Nmd3 in a Crm1-dependent manner. Recently the general mRNA export factor, the Mtr2/Mex67 heterodimer, was shown to act as an export receptor in parallel with Crm1. These observations raise the possibility that nuclear export of the 60S subunit in Saccharomyces cerevisiae requires multiple export receptors. Here, we show that the previously characterized 60S subunit biogenesis factor, Arx1, also acts as an export receptor for the 60S subunit. We found that deletion of ARX1 was synthetic lethal with nmd3 and mtr2 mutants and was synthetic sick with several nucleoporin mutants. Deletion of ARX1 led to accumulation of pre-60S particles in the nucleus that were enriched for Nmd3, Crm1, Mex67, and Mtr2, suggesting that in the absence of Arx1, 60S export is impaired even though the subunit is loaded with export receptors. Finally, Arx1 interacted with several nucleoporins in yeast two-hybrid as well as in vitro assays. These results show that Arx1 can directly bridge the interaction between the pre-60S particle and the NPC and thus is a third export receptor for the 60S subunit in yeast.

Download Full-text