What type of cell is rough er

The rough endoplasmic reticulum is made up of membranes which have ribosomes attached to it. This gives it the rough appearance. This is contrary to the smooth endoplasmic reticulum which does not have these membrane bound ribosomes.

This means that the rough endoplasmic reticulum is responsible for the production of polypeptides that are about to be taken through the membrane to be post-modified.

Rough endoplasmic reticulum is an organelle found in eukaryotic cells. Its main function is to produce proteins. It is made up of cisternae, tubules and vesicles. The protein might work well without the amino acid but the very exacting service provided by the quality control section spots the error and rejects the protein retaining it in the lumen of the rough ER. In this case the customer the person with cystic fibrosis loses out completely due to high standards when a slightly poorer product would have been better than no product at all.

They are conveyed in vesicles or possibly directly between the ER and Golgi surfaces. It is found fairly evenly distributed throughout the cytoplasm. Smooth ER is devoted almost exclusively to the manufacture of lipids and in some cases to the metabolism of them and associated products.

In liver cells for example smooth ER enables glycogen that is stored as granules on the external surface of smooth ER to be broken down to glucose.

Smooth ER is also involved in the production of steroid hormones in the adrenal cortex and endocrine glands. Smooth ER — the detox stop Smooth ER also plays a large part in detoxifying a number of organic chemicals converting them to safer water-soluble products. Large amounts of smooth ER are found in liver cells where one of its main functions is to detoxify products of natural metabolism and to endeavour to detoxify overloads of ethanol derived from excess alcoholic drinking and also barbiturates from drug overdose.

To assist with this, smooth ER can double its surface area within a few days, returning to its normal size when the assault has subsided. The contraction of muscle cells is triggered by the orderly release of calcium ions.

These ions are released from the smooth endoplasmic reticulum. In the early phase of translation, a signal peptide is synthesized i. The signal is an indication that the protein is for further processing in the ER. When this signal is recognized by a signal recognition particle the ribosome translating the protein docks to the endoplasmic reticulum via the translocon.

The ribosome, then, returns back to the translation of the protein. The chain continues to grow as the mRNA transcript is translated through the docked ribosome. The chain eventually makes its way into the ER through the translocon that spans across the ER membranes. The signal peptide is removed by a signal peptidase in the lumen of the ER. The nascent protein is folded in the ER by the chaperone proteins e.

The properly-folded protein is then packed into a transport vesicle to be shuttled to the Golgi apparatus where it would undergo maturation for transport along the cytoskeleton to other cytoplasmic organelles like lysosomes and peroxisomes or for secretion out of the cell. Some of the proteins synthesized inside the ER will be retained, such as those that become part of the ER membrane.

Those that are retained in the ER have a retention motif, e. An unfolded or misfolded protein triggers an endoplasmic reticulum stress response. This happens when certain disturbances occur, such as disturbances in the redox regulation, calcium regulation, viral infection, and glucose deprivation.

A distinctive feature of a misfolded protein is the lack of glucose residues, which are attached via N-linked glycosylation. A heat shock protein glucose regulate protein 78 may bind to the hydrophobic residues of the misfolded protein to prevent its transit.

If protein misfolding continues, the protein is headed towards degradation to prevent it from aggregating with other misfolded proteins. Overexpression of some reticulon isoforms leads to formation of long ER tubules at the expense of sheets [ 58 ]. In turn, depletion of reticulons, and hence the ability to bend membranes, leads to a reduction in the number of ER tubules, leading to an expansion of peripheral sheets [ 57 , 59 , 60 ].

Therefore, the level of reticulons within a cell determines the abundance and fine structure of ER tubules. Reticulons do not act alone in shaping ER tubules. Atlastins, members of the dynamin-like GTPase family, mediate these homotypic fusion events. Depletion by RNAi or expression of dominant-negative atlastin in cells results in a lack of fusion events leading to an abundance of long, unbranched tubules [ 61 ].

When a dominant-negative cytoplasmic fragment from Xenopus , which contains the GTPase domain but lacks the transmembrane domain and cytoplasmic tail [ 64 ], are introduced into Xenopus interphase extracts ER network formation was blocked [ 65 ]. Comparable point mutations that prevent dimerization of the cytoplasmic fragment of human atlastin [ 66 ] were made in the Xenopus cytoplasmic atlastin protein, added into interphase extract and had no effect on ER network formation [ 65 ].

Furthermore, antibodies directed against atlastin inhibit ER network formation when introduced into Xenopus egg extracts [ 61 ].

In Drosophila , atlastin depletion leads to ER fragmentation and purified atlastin is sufficient to catalyze GTP-dependent fusion of proteoliposomes [ 64 , 66 , 67 ].

Therefore, studies from multiple organisms, extracts and purified components indicate that atlastin is likely required for catalyzing homotypic vesicle fusion between ER membranes, which is important for proper network formation. Recently, a few new key players have been identified that are involved in ER dynamics. Work using purified ER vesicles derived from Xenopus eggs has demonstrated that GTP is required for homotypic ER vesicle fusion in the absence of cytosolic factors [ 57 , 68 ].

Previous studies indicated that GTPases are required for ER fusion events [ 69 , 70 ], and a recent study utilized a proteomics approach to identify Rab10 as a factor required for ER assembly [ 71 ]. Knock-down of Rab10, or overexpression of a GDP-locked dominant-negative point mutant, in cultured human cells caused an increase in ER sheets and a decrease in tubules [ 71 ].

ER—ER fusion events occurred at regions where Rab10 was enriched. It is currently not clear what role Rab10 plays in the ER vesicle fusion reaction or how homotypic ER vesicle fusions are coupled to lipid synthesis. Depletion of Rab18 leads to a phenotype similar to that observed following Rab10 inhibition [ 72 ]. Additionally, when Rab10 is depleted, Rab18 redistributes to peripheral sheets [ 72 ].

Therefore, it appears that depletion of either Rab10 or Rab18 prevents the stabilization of ER tubule fusion, reducing the density of tubules resulting in an increase in ER sheets. In addition to the role RAB-5 plays in peripheral ER formation, kinetics of nuclear envelope disassembly is affected in these mutants [ 70 ]. In addition to GTPases that may play a direct role in homotypic membrane fusion of vesicles, recent work has demonstrated a role for lipid synthesizing enzymes in controlling the shape and organization of the ER.

Inhibition of C-terminal domain CTD nuclear envelope phosphatase-1 CNEP-1 , which is enriched on the nuclear envelope and promotes the synthesis of membrane phospholipids, led to the appearance of ectopic sheets that encased the nuclear envelope, interfering with nuclear envelope breakdown [ 74 ]. These results reflect the interconnected network of proteins and functions that play a role in shaping the structures of the ER.

The ER is a very dynamic network that is constantly undergoing rearrangements and remodeling [ 75 ]. ER tubules are continually fusing and branching resulting in the creation of new three-way junctions. In a competing process, junction sliding and tubule ring closure leads to loss of three-way junctions and the characteristic polygonal structure [ 76 ].

Very little is known about the complexes controlling this process, but it was recently discovered that Lunapark Lnp1 localizes to and stabilizes three-way junctions [ 77 , 78 ]. Lnp1 binds to reticulons and Yop1, and localization of Lnp1 to junctions is regulated by Sey1p, the yeast homolog of atlastin [ 78 ].

Loss of Lnp1 leads to a collapsed and densely reticulated ER network in yeast and human cultured cells [ 77 , 78 ], though only half of the junctions are bound to Lnp1 [ 77 ], which reflects the fluidity of the ER network. If Lnp1 is overexpressed, the protein localizes to the peripheral ER and induces the formation of a large polygonal tubular network [ 79 ].

Additionally, formation of this network was inhibited by Lnp1 mutations that blocked N -myristoylation [ 79 ], an attachment of myristic acid a carbon saturated fatty acid , indicating that this modification plays a critical role in Lnp1-induced effects on ER morphology.

N -myristoylation is not required for membrane translocation, topology formation, or protein localization to the ER but may play a role in protein—protein or protein-lipid interactions that are required for morphological changes in the ER, though the exact molecular mechanism of action remains to be elucidated [ 79 ].

The actual mechanism for Lnp1-mediated stabilization of three-way junctions is unknown, though recent studies and insights from the structure and domains within the protein shed light on how Lnp1 stabilizes junctions [ 77 , 78 ]. First, Lnp1 contains two transmembrane domains as well as a zinc finger domain, which is located on the cytoplasmic face of the ER membrane [ 77 ]. When cysteines were mutated within the zinc finger domain, the polygons became smaller and regions lacking cortical ER were more apparent as the number of cysteines mutated increased [ 78 ].

Therefore, mutations in the zinc finger domain may affect protein—protein interactions, complex formation or interfere with the distribution of resident lipids on the cytoplasmic face of the membrane causing deleterious effects on junction stabilization. In addition, the transmembrane domains may be acting as an inverted wedge, adding to the local negative curvature characteristic of three-way junctions [ 77 ], and acting opposite to the positive curvature promoted by reticulons.

Another possibility is that multiple Lnp1 proteins may also act cooperatively together to stabilize the junction, or Lnp1 may be acting transiently to stabilize or modify lipids or other proteins at junctions [ 77 ].

In addition to proteins that regulate membrane structure and dynamics, there is accumulating evidence that changing the nucleic acid content of the ER can also impact ER shape. Early experiments showed that brief treatment of tissue culture cells with the translation inhibitor puromycin, which dissociates mRNA:ribosome complexes, leads to loss of ribosomes from the ER and a loss of ER sheets [ 51 , 80 ].

Depletion of XendoU leads to the formation of long, unbranched tubules in Xenopus leavis egg extract, and rescue of this phenotype requires intact catalytic activity of the protein, indicating that the nuclease function is critical to proper ER network formation [ 82 ].

Furthermore, antibody addition to purified vesicles leads to a block in network formation, demonstrating that XendoU acts on the surface of ER membranes to regulate ER structure [ 82 ]. Depletion of XendoU also leads to a delay in replication and nuclear envelope closure [ 82 ], and BAPTA blocks nuclear envelope formation in Xenopus egg extract reconstitution experiments [ 85 ].

Upon vesicle fusion it was found that RNAs were degraded and released from the surface of membranes, suggesting that XendoU acts to degrade these RNAs, as well as release proteins, to clear patches of membrane to allow for vesicle formation leading to network formation [ 82 ].

Interestingly, when purified vesicles were treated with increasing concentrations of RNaseA and subjected to the same assay, an increasingly aberrant network formed with large vesicles that were unable to fuse [ 82 ].

Results from in vitro studies indicate that XendoU is activated on membranes in coordination with calcium release to locally degrade RNAs and clear patches of membranes leading to fusion in a controlled manner to fine tune network formation. Lastly, similar to other proteins that play a role in tubule formation, knock-down of the human homolog EndoU in cultured human cells leads to an expansion of sheets [ 82 ].

Additionally, rescue of the expanded sheet phenotype depended on intact catalytic function as observed with recombinant protein in the extract system. Therefore, XendoU is an example of a protein that is activated in response to cellular cues to regulate proper ER formation, and further studies may reveal additional proteins that are regulated in this manner to fine tune organelle structure. We have considered how tubules are formed and maintained, which leads the discussion to sheets, the other peripheral ER structure.

First, we must consider how sheets are formed. Several mechanisms have been proposed, including the idea that integral membrane proteins can span the intraluminal space and form bridges, connecting the lipid bilayers [ 51 , 86 , 87 ]. These proteins may either stabilize the structure or define the distance between the two lipid layers based on the size of the proteins. Additionally, these proteins or protein complexes may form a scaffold that aids in the stabilization of the sheets or bring the two lipid membranes in closer proximity [ 86 ].

Several proteins including Climp63, p and kinectin have been implicated in the generation, maintenance and stabilization of ER sheets [ 51 ]. In addition to highly enriched membrane proteins and core components of the translocon, Climp63, a coiled—coiled protein with a single transmembrane domain, was identified along with kinectin and p in a mass spectrometry screen for abundant integral ER membrane proteins [ 51 ].

Through various techniques and in various cell types Climp63 was shown to be a highly abundant protein [ 88 — 90 ] that localizes to perinuclear ER and is absent from the nuclear envelope [ 91 , 92 ].

Very stable oligomers of Climp63 can form, restricting mobility of the protein along the membrane, promoting localization to the rough ER [ 92 ]. Overexpression of Climp63 leads to a massive proliferation of ER sheets while reduction in expression surprisingly does not lead to loss of sheets but instead a decrease in the distance between sheets [ 51 ]. Moreover, these sheets are spread diffusely throughout the cytoplasm, reminiscent of the phenotype of cells treated with the translation inhibitor puromycin [ 51 ].

This is interesting as the core components of the translocon, the protein channel that interacts with ribosomes and is responsible for translocating nascent peptides into the ER or anchoring transmembrane segments of newly synthesized proteins, were found to be enriched on sheets [ 93 ]. Therefore, these results suggest that the role of Climp63 in formation of sheets is likely to involve additional factors and acts as a part of an elaborate regulatory network that balances the production of sheets and tubules.

It is clear that proteins involved in the promotion, maintenance or stabilization of peripheral ER structures function through interactions with additional proteins or structures, and these interactions are key to proper formation of the ER network. Interestingly, several of the proteins discussed above have been shown to interact with microtubules, including Climp63 [ 91 ], p [ 94 ], kinectin [ 95 ] and STIM1 discussed below.

One important interaction discussed below is with microtubules. The ER network exhibits several dynamic interactions with microtubules that are important for determining the distribution of the ER within the cell. The two main types of interactions between the ER and microtubules are Tip Attachment Complexes TACs and sliding along preformed microtubules by the action of kinesin and dynein motors [ 96 — ].

In cultured cells treated with nocodazole to depolymerize microtubules, the ER retracts from the periphery [ ], though the retraction does not occur immediately. Further investigation revealed that sliding events occurred mainly on a small subset of microtubules, modified by acetylation, that are more resistant to nocodazole treatment [ 76 ]. Furthermore, ER tubules can form in the absence of microtubules [ 57 , 65 , 68 ], raising many questions and leading several groups to study the interaction between ER and microtubules more in-depth.

In the past 10 years we have learned a great deal about what proteins are responsible for the intrinsic shape of the ER and how these proteins are connected to specific ER subdomains. However, we know very little about how cellular signals communicate with ER shaping proteins to change the shape of the ER in response to cellular signals.

During mitosis many cellular structures are dramatically remodeled to facilitate chromosome segregation. One of the most dramatic examples is changes to the microtubule cytoskeleton that occur as a result of increased microtubule dynamics caused by the action of cyclin-dependent kinases.

The increase in microtubule dynamics during mitosis is important for the bipolar attachment of chromosomes to the mitotic spindle and accurate segregation to daughter cells during anaphase [ ]. In addition to changes to the microtubule cytoskeleton, essentially all organelles change shape and function during mitosis to facilitate accurate organelle inheritance and orderly chromosome segregation.

The ER undergoes dramatic shape changes during mitosis and recent studies are beginning to uncover the mechanisms linked to these structural changes. In organisms with an open mitosis the nuclear envelope breaks down at the onset of mitosis to allow free exchange between the nucleus and cytoplasm.

Nuclear envelope breakdown NEBD is a carefully orchestrated process that begins during mitotic prophase [ ]. During prophase components of the nuclear pore dissociate from the pore, the nuclear lamina depolymerizes, and the membrane-bound proteins of the nuclear envelope retract into the general ER.

These events free the chromosomes of nuclear lamina and membranes to facilitate chromosome condensation and segregation.

In general, the events of nuclear envelope breakdown are thought to be driven by the phosphorylation of components of the NE during mitosis by various mitotic kinases, especially cyclinB:cdk1, although many molecular details are still unclear. Concomitant with changes that occur to the nuclear envelope during NEBD the ER also begins to undergo dramatic shape changes. Changes in ER shape during mitosis have been studied in many different organisms by both light and electron microscope and these studies have resulted in a conflicting series of reports about the shape of the ER during mitosis.

However, during the last few years a consensus has begun to emerge that the mitotic ER is primarily composed of sheets. Early studies using live cell microscopy in both Drosophila and C.

Additionally, work using thin section transmission EM in HeLa cells also concluded that the majority of the ER was present in sheets throughout mitosis [ ]. However, two studies in a variety of mammalian tissue culture cells [ 80 , ] have used both live cell microscopy and electron microscopy to suggest that the ER is primarily tubular during mitosis, and two additional studies [ 60 , ] also suggested that the ER remained tubular during mitosis and further suggested that end-on binding of ER tubules to chromatin during mitosis initiates nuclear envelope reassembly at the end of mitosis.

One potential difficulty in interpreting the shape of the mitotic ER is that most cells round up during mitosis which can make acquisition of light and electron microscopy images difficult and require laborious reconstruction of the images into a three dimensional model. In addition, the mitotic ER is highly dynamic, which can complicate acquisition of live cell images during mitosis.

To address these questions a series of recent studies have used both high-resolution, high-speed live cell microscopy and high-resolution EM to demonstrate that the ER is almost exclusively composed of sheets during mitosis [ , ]. In addition, these studies demonstrate that the nuclear envelope reforms through the docking of ER sheets onto regions of chromatin that are isolated from spindle microtubules [ ].

Finally, to circumvent many or the problems associated with imaging large, three dimensional cells during mitosis a recent study has examined the structure of the ER in vitro using ER reconstituted from Xenopus egg extracts [ 65 ]. This study convincingly demonstrated that ER formed in mitotic extracts is primarily composed of sheets while interphase ER is primarily composed of tubules. In addition, the authors demonstrated that active cyclinB:cdk1 was sufficient to convert a tubular ER into a primarily sheet based ER.

Taken together all of these studies present conflicting views of the shape of the ER during mitosis, but a consensus is emerging from a wide variety of organisms that the mitotic ER is primarily composed of sheets and that the shape changes in the ER are related to changes in cyclin:cdk activity. In addition to changes in the gross morphology of the ER during mitosis there are also dramatic changes in the distribution of proteins throughout the ER.

During interphase the ER is organized into distinct domains with certain proteins defining different domains. For example, the tubule-shaping reticulon protein Rtn4 is exclusively present in the peripheral ER and excluded from the nuclear envelope [ 57 , 60 , ]. However, during mitosis the NE retracts into the ER and there is nearly complete mixing of the specialized ER-shaping proteins [ 60 , ]. At the end of mitosis proteins that define the NE and peripheral ER are rapidly resorted such that they reestablish their characteristic interphase organization [ 60 , ].

In addition, it has been shown that overexpression of Rtn4 or knockdown of three reticulons Rtn1, Rtn3, Rtn4 can either slow or speed the rate of NE reassembly at the end of mitosis, although the mechanism through which these proteins affect NE formation is currently unknown. These studies highlight the massive reorganization that takes place in the ER during mitosis and suggests that different expression levels of specific ER shaping proteins can control ER reorganization during mitosis.

However, we know very little about how various ER shaping proteins are resorted to specific domains at the end of mitosis. Two very recent studies [ , ] have begun to provide insight into the specialized processes that regulate nuclear envelope reformation at the end of mitosis. Both of these studies identified a transient localization of the ESCRT-III complex to the surface of chromatin during late anaphase when the nuclear envelope is beginning to reform.

ESCRT-III is best known for its role in the formation of multivesicular bodies during endocytosis, but also has well-documented roles in cytokinesis and viral budding from the plasma membrane [ ]. Additionally, interactions with the microtubule severing enzyme spastin and the ubiquitin recognition factor UFD1 are important for nuclear envelope reformation.

The redistribution of ER shaping proteins during mitosis suggests that the fundamental activities of some of these proteins are modified during mitosis. For example, the mitotic ER is composed of primarily sheets, yet Rtn4, which promotes tubule formation [ 57 ], is distributed throughout the ER [ 60 , ].

This result suggests that the tubule-promoting activity of Rtn4 may be modified during mitosis to facilitate the tubule-to-sheet transition observed during mitosis. Inspection of large-scale phospho-proteomics studies reveals that a large number of ER-shaping proteins have identified mitosis-specific phosphorylation sites [ — ].

Although none of the phosphorylation sites identified in these large-scale screens has been studied in detail their presence and specificity to mitosis suggests that these are likely to be involved in reshaping the ER during mitosis. In support of the hypothesis that mitosis-specific phosphorylation of ER-shaping proteins regulates ER remodeling during mitosis two studies have examined this phenomenon in detail. A study of the ER sheet promoting protein Climp63 [ 51 ] has demonstrated mitosis-specific phosphorylation on three N-terminal residues [ ].

Phosphorylation of Climp63 blocks the interaction of Climp63 with microtubules. Additionally, phosphomimetic mutants blocked the interaction of the ER with microtubules during interphase and resulted in an ER composed primarily of sheets, while nonphosphorylatable mutants tethered the ER to microtubules and resulted in an extremely distorted ER.

These results suggest that mitotic phosphorylation of Climp63 likely blocks the interaction of the ER with microtubules and could be an important step in the tubule-to-sheet transition that occurs during mitosis. A second study examined the interaction of the ER with growing microtubule plus ends during mitosis.

However, during mitosis the ER is excluded from the mitotic spindle and does not exhibit plus tip growth events. A recent study [ ] has demonstrated that STIM1 is specifically phosphorylated during mitosis to control the interaction of the ER with microtubules.

Clearly much more work remains before we have a clear understanding of how cell cycle signaling cascades contribute to reshaping of the mitotic ER. While the above studies demonstrated that phosphorylation of key proteins that link the ER to the microtubule cytoskeleton is important for excluding the ER from the spindle during mitosis a recent study demonstrated the importance of an interaction of the ER with microtubules for clearing the ER from mitotic chromatin.

During mitosis the nuclear envelope is absorbed into the ER and is cleared from the surface of the chromatin, however little is known about the mechanisms that regulate ER removal from the chromatin.

Taken together these three studies demonstrate that interaction of the ER with microtubules is a major mechanism that contributes to shape rearrangement during mitosis and that ER:microtubule interactions are regulated by mitotic phosphorylation.