Analysis of processing and polyadenylation signals of the hepatitis B virus surface antigen gene by using simian virus 40-hepatitis B virus chimeric plasmids. Identification of a sequence element on the 3' side of AAUAAA which is necessary for simian virus 40 late mRNA 3'-end processing. Sadofsky M, Connelly S, Manley JL, Alwine JC.Sequences on the 3' side of hexanucleotide AAUAAA affect efficiency of cleavage at the polyadenylation site. 3' non-coding region sequences in eukaryotic messenger RNA. Regulation of adenovirus-2 gene expression at the level of transcriptional termination and RNA processing. The pathway of eukaryotic mRNA formation. Adenovirus gene expression: control at multiple steps of mRNA biogenesis. Accurate cleavage and polyadenylation of exogenous RNA substrate. Inhibition of RNA cleavage but not polyadenylation by a point mutation in mRNA 3' consensus sequence AAUAAA. Montell C, Fisher EF, Caruthers MH, Berk AJ.Targeted random mutagenesis: the use of ambiguously synthesized oligonucleotides to mutagenize sequences immediately 5' of an ATG initiation codon. The consensus sequence YGTGTTYY located downstream from the AATAAA signal is required for efficient formation of mRNA 3' termini. McLauchlan J, Gaffney D, Whitton JL, Clements JB.Requirement of a downstream sequence for generation of a poly(A) addition site. McDevitt MA, Imperiale MJ, Ali H, Nevins JR.Mutations downstream of the polyadenylation site of a Xenopus beta-globin mRNA affect the position but not the efficiency of 3' processing. Mason PJ, Elkington JA, Lloyd MM, Jones MB, Williams JG.An enhancer-like element in the adenovirus E2 promoter contains sequences essential for uninduced and E1A-induced transcription. A small nuclear ribonucleoprotein associates with the AAUAAA polyadenylation signal in vitro. Poly(A) site cleavage in a HeLa nuclear extract is dependent on downstream sequences. Definition of essential sequences and functional equivalence of elements downstream of the adenovirus E2A and the early simian virus 40 polyadenylation sites. Hart RP, McDevitt MA, Ali H, Nevins JR. Characteristics of a human cell line transformed by DNA from human adenovirus type 5. Graham FL, Smiley J, Russell WC, Nairn R.A sequence downstream of AAUAAA is required for rabbit beta-globin mRNA 3'-end formation. The sequence 5'-AAUAAA-3'forms parts of the recognition site for polyadenylation of late SV40 mRNAs. A sequence downstream of A-A-U-A-A-A is required for formation of simian virus 40 late mRNA 3' termini in frog oocytes. Identification of sequences in the herpes simplex virus thymidine kinase gene required for efficient processing and polyadenylation. Transcription termination and 3' processing: the end is in site! Cell. Calcitonin/calcitonin gene-related peptide transcription unit: tissue-specific expression involves selective use of alternative polyadenylation sites. Links to PubMed are also available for Selected References. Get a printable copy (PDF file) of the complete article (1.6M), or click on a page image below to browse page by page. Full textįull text is available as a scanned copy of the original print version. This observation, along with the fact that the two sequences are clearly different, indicates that there are at least two distinct genetic elements that direct efficient cleavage at the poly(A) site. There were differences, however, in the effect of spacing on the function of the two elements. The position of the downstream elements relative to the AAUAAA and cleavage site was found to be critical since moving either the E2 element or the SV40 element an additional 40 nucleotides downstream abolished function. Thus, it is concluded that a defined downstream sequence of limited complexity is important for efficient processing of the primary transcript at the poly(A) site. Certain single base changes drastically altered poly(A) site function. A series of base substitution mutants were generated in each downstream sequence. Inversion of the sequence completely abolished poly(A) site function. Chemically synthesized oligonucleotides of sequence from the early SV40 and the adenovirus E2A poly(A) sites were able to restore efficient cleavage to a deleted SV40 poly(A) site. Two downstream regions were analyzed in an attempt to accurately locate and define the critical sequences. Several recent studies have shown that a functional poly(A) site consists of both an AAUAAA element as well as sequences downstream of the cleavage site.
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Speaking of game world, I found it to be rather immersive although the graphics are neither stylish nor realistic enough to truly stand out. Not only that even though you make your own character, they even say things mid-gameplay which makes the world feel all the more real as the things they say always match their current situation. With that in mind, I was very impressed with Remnant: From the Ashes' cast of NPCs who range from sarcastic introverts to helpful fellows and suspicious drifters to wise old folks. Most Souls-like games lack a sense of personality as they generally try to create mysterious atmospheres while leaving your character as a blank slate. Then, simply blow everything else away with a shotgun. Have a hulking monster on your tail? Smash him up with your hammer. Being swarmed by ankle biters? Rapid-fire shoot them with a pistol. Anyway, you basically equip 2 firearms as well as a melee weapon and can quickly alternate between the 3 depending on your current situation. For starters, it features both long-range and melee combat which I guess you could compare to Bloodborne although that focuses much more on melee than Remnant: From the Ashes does. To my surprise, Remnant: From the Ashes doesn't feel as Souls-like as most of its brethren so right off the bat, I knew I was in for a unique adventure. □ Prepare to see plenty of dilapidated buildings and withering treesĬoming fresh from playing the incredible Nioh 2, I wanted to sink my teeth into yet another Dark Souls style ARPG. has been obsessively gaming since the late '80s and is just as passionate about video games in 2023. Last year's action RPG hit Remnant: From the Ashes now has a swanky new physical release so I figured it's about time to check it out. 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If you have questions about the status of a GST/HST objection that you’ve filed and you don’t have your Appeals Officer’s contact information, you can call the following: The appeals process to have a CRA officer resolve your GST/HST objection can take anywhere from 124 days to over 500 days (as of January 2021) - it depends on the level of complexity of your GST/HST objection. Register for a business number and any program account such as the GST/HST. Report a nil remittance for non-resident withholding tax. T4, T5, T3 Filing information returns electronically. Get help with the Corporation Internet Filing service.ĭiscuss payment arrangements to repay a GST/HST debt (paying over several months).ĭiscuss payment arrangements to repay a payroll tax debt (paying over several months).ĭiscuss payment arrangements to repay a corporate income tax debt (paying over several months). Get help with filing your T2 corporation income tax return electronically. Have a payment made in the wrong business account transferred to another account.īusiness tax for corporations, sole proprietorships, partnerships, non-resident corporations, and corporation accounts. Yukon, Northwest Territories and Nunavut: Help with My Business Account and Represent a Client. Let’s review the telephone numbers for you to reach a perky CRA agent. Request the agent ID number and record details of your conversation either on a note-taking device or on paper.Īlthough there are some spooky tax shenanigans that can happen, your chances of getting the culprit fixed depend on your attention to the telephone number you call. At the beginning of the call, inform the agent of a call back number they can reach you at if for some reason you become disconnected. Use a speakerphone to ensure you can have your hands free to review files for questions asked by the CRA agent or log in to your client’s portals to confirm information. □ Fact: CRA agents abide by a Taxpayer Bill of Rights that must be followed during every taxpayer interaction. Be kind and courteous to our fellow public service employees. Paper and pen to take notes or a note-taking device.ĬRA agents are quite nice and respectful. Inquiry details, such as the latest notice of assessment, return or other tax documents. Your client’s business information, including: You will also want to have all the following on hand: Plan your call time during normal business hours and non-public holidays.Ĭheck the anticipated wait times to speak to a CRA agent.Įnsure that you have the authorization to speak to the CRA on behalf of your client. Dos and Don’ts when calling the CRA.Īs they say, “A little preparation goes a long way.” In the spirit of making things a little less taxing, I’m sharing my tips and top phone numbers to spend less time on hold and put more time back in your business. Emails are a frequent way to communicate on a timely basis, but the CRA does not send or receive emails relating to business inquiries. In our fast-paced world, sometimes picking up the phone is the best way to ensure proper communication and get a problem resolved for our clients.Īlthough you may not have time for a friendly chat, our friends at the Canada Revenue Agency (CRA) do. |
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