• Description
  • Highlights
  • Detected Modifications
  • Workflow
  • Data Analysis
Service NamePrice
LC-MS tRNA Modification Analysis Service

tRNAs are the fundamental component of mRNA decoding and protein translation. tRNAs undergo by far the greatest number of and the most chemically diverse post-transcriptional modifications. These modifications are critical for all core aspects of tRNA function, such as folding, stability and decoding[1]. Typically, modifications in the main body of tRNA are crucial for tRNA structure folding, stability, rigidity and flexibility, whereas modifications in the anticodon loop affect decoding by open loop structure, codon-anticodon pairing, wobbling, and preventing translational frameshifts. Additionally, modified nucleosides serve as identity determinants for aminoacyl-tRNA synthetase (AARS) for extra amino acid recognition accuracy[2]. In general, hypomodified tRNAs are targeted for degradation. Studying tRNA modification is perhaps just as important as tRNA expression profiling.

Defects in tRNA modifications and modification enzymes are linked with human diseases such as cancers, diabetes, neurological syndromes, cardiac conditions, and mitochondrial-linked disorders (Fig. 1)[3]. Analysis of tRNA modification profiles is key to establish the link with the disease, tRNA modification enzymes, and tRNA molecular functioning.

Arraystar LC-MS tRNA Modification Analysis Service offers the sample-to-data solution for simultaneous profiling of 36 nucleosides modifications important to tRNA, using total RNA as the starting material. The service includes tRNA isolation from the total RNA, complete hydrolysis, and dephosphorylation to prepare single nucleosides. The state of the art, ultra high performance LC-MS system delivers a new level of sensitivity, precision, accuracy, dynamic range, and robustness of the quantification results.

1

Figure 1. tRNA modifications in human diseases

References
1. Kirchner S. and Z. Ignatova (2015) "Emerging roles of tRNA in adaptive translation, sig-nalling dynamics and disease." Nat. Rev. Genet. 16(2):98-112 [PMID: 25534324]
2. El Yacoubi B. et al. (2012) "Biosynthesis and function of posttranscriptional modifications of transfer RNAs." Annu. Rev. Genet. 46:69-95 [PMID: 22905870]
3. Torres A.G. et al. (2014) "Role of tRNA modifications in human diseases." Trends Mol Med 20(6):306-14 [PMID: 24581449]

 

• Quantitative analysis of complex tRNA modifications from total RNA samples.
• Full service sample-to-data — from sample QC, tRNA isolation, nucleoside analyte prepara-tion, LC-MS/MS data acquisition, analysis to report.
• High Performance— Highly optimized experimental procedures, state-of-the-art LC-MS sys-tem, expertise in operation.
• Best analytical coverage—Simultaneous profiling of 36 nucleoside modifications in tRNA.

 

Table1. Nucleoside modifications profiled by Arraystar LC-MS tRNA Modification Analysis

Number

Nucleoside

Symbol

Number

Nucleoside

Symbol

01

3'-O-methyladenosine

3'-OMeA

19

1-methylguanosine

m1G

02

2'-O-methylcytidine

Cm

20

7-methylguanosine

m7G

03

5-methylcytidine

m5C

21

N2-methylguanosine

m2G

04

N6-isopentenyladenosine

i6A

22

3'-O-methylinosine

3'-OMeI

05

5,2'-O-dimethylcytidine

m5Cm

23

2-thiouridine

s2U

06

1-methyladenosine

m1A

24

4-thiouridine

s4U

07

2-thiocytidine

s2C

25

5-methyluridine

m5U

08

N2,N2,7-trimethylguanosine

m2,2,7G

26

N4-acetylcytidine

ac4C

09

N4-acetyl-2'-O-methylcytidine

ac4Cm

27

3'-O-methyluridine

3'-OMeU

10

N6-methyladenosine

m6A

28

5-methyl-2-thiouridine

m5s2U

11

3-methylcytidine

m3C

29

5-methoxyuridine

mo5U

12

3'-O-methylcytidine

3'-OMeC

30

pseudouridine

13

2'-O-methyladenosine

Am

31

2'-O-methylinosine

Im

14

N2,N2-dimethylguanosine

m22G

32

3-methyluridine

m3U

15

5'-O-methylthymidine

5'-OMeT

33

3'-O-methylguanosine

3'-OMeG

16

2'-O-methyluridine

Um

34

1-methylpseudouridine

17

inosine

I

35

5-hydroxymethylcytidine

hm5C

18

2'-O-methylguanosine

Gm

36

5,2'-O-dimethyluridine

m5Um

2

Workflow of Arraystar LC-MS tRNA Modification Analysis.

• Raw and normalized peak data

3

• Total Ion Current chromatogram of nucleosides

4

• Differential modification of nucleosides among samples

5

References

1. Kirchner S, Ignatova Z. Emerging roles of tRNA in adaptive translation, signalling dynamics and disease. Nature reviews Genetics 2015;16:98-112.
2. El Yacoubi B, Bailly M, de Crecy-Lagard V. Biosynthesis and function of posttranscriptional modifications of transfer RNAs. Annual review of genetics 2012;46:69-95.
3. Torres AG, Batlle E, Ribas de Pouplana L. Role of tRNA modifications in human diseases. Trends in molecular medicine 2014;20:306-14.