Molecular Biology Flashcards
What type of RNAs code for proteins?
mRNA
What type of RNA forms the basic structure of the ribosome and catalyzes protein synthesis?
rRNA (ribosomal)
What type of RNA is central to protein synthesis as an adaptor between mRNA and amino acid?
tRNA (transfer)
What type of RNA functions in a variety of nuclear processes, including the splicing of pre-mRNA?
snRNA (small nuclear)
What type of RNA helps to process and chemical modify rRNAs?
snoRNA (Small nucleolar)
What type of RNA regulates gene expression by blocking translation of specific mRNAs and causes their degradation?
miRNA (micro)
What type of RNA turns off gene expression by directing the degradation of selective mRNAs and the establishment of compact chromatin structures?
siRNA (small interfering)
Prokaryotic transcription requires recruitment of _______ factor to the RNA polymerase. In order for transcription to continue and elongation to occur, this factor must be released.
Sigma
Initiation of eukaryotic transcription begins with the binding of _______ to the ______ box
TFIID
TATA
[the subunit of TFIID that recognizes TATA is called TATA-box-binding protein or TBP]
During initiation of eukaryotic transcription, once TFIID has bound the TATA box, what is the next factor to be recruited? This is followed by what other factors?
TFIIB, followed by TFIIF, RNA polymerase II, TFIIE, and TFIIH
Which of the initiation factors in eukaryotic transcription serves to unwind the DNA double helix and expose the DNA template strand?
TFIIH
During initiation of eukaryotic transcription, phosphorylation of _______ by TFIIH causes the polymerase to leave the promoter and begin elongation
CTD
In short, the most important factor for initiation in prokaryotic transcription is _______, while in eukaryotic transcription it is the ______ factors.
Sigma
TFII
What 3 methods of processing must RNA go through in eukaryotics prior to translation?
RNA capping at 5’ end (with 7-methyl guanosine)
RNA splicing (intron removal)
Polyadenylation at 3’ end (addition of poly A tail)
Each group of 3 consecutive nucleotides in RNA is called a codon. Each codon specifies either one amino acid or a stop in the translation process. A total of ____ triplet codons code for the _____ known amino acids, thus the code is _____________
61
20
Degenerate
What is the start codon of the genetic code?
Met = AUG
What are the 3 possible stop codons in the genetic code?
UAA
UAG
UGA
The ____________ hypothesis postulates that the genetic code is read by molecules that can recognize a codon and carry the corresponding amino acid.
Adaptor
What serves as an adaptor that binds to a specific codon and brings with it an amino acid for incorporation into the polypeptide chain?
tRNA
tRNA has a ____________ secondary structure with 4 short double-helical segments. Two regions of unpaired nucleotides are crucial to the functions of tRNA: the _____________ loop, which is a set of 3 consecutive nucleotides that pair with a complementary codon in an mRNA molecule, and the ________ __________ region which binds the amino acid that matches the corresponding codon.
Cloverleaf
Anticodon
3’ CCA terminal
The genetic code is translated by 2 adaptors: how does the correct tRNA with the correct amino acid come to the mRNA?
Aminoacyl tRNA synthetase has a binding site for both amino acid and codon
During translation, what enzyme catalyzes the translocation of 2 complete subunits to the A site?
Peptidyl transferase
What are the 3 ribosomal sites E, P, and A for during translation?
A = incoming tRNA with new amino acid
P = for polypeptide chain
E = exit
During elongation, two things are occuring simultaneously, one is the growth of the amino acid chain, what is the other?
Proofreading, as incorrectly base-paired tRNAs preferentially dissociate
What is unique about stop codons?
They are not recognized by any tRNA and do not code for any amino acid
What type of protein recognizes stop codons and what do they do?
Release Factors (RFs), which promote release of completed proteins from tRNAs by binding to the A site of the ribosome containing the stop codon and catalyzing the addition of water instead of an amino acid
What are the 3 general types of protein transport?
Gated transport
Transmembrane transport
Vesicular transport
Protein transfer/transport to various compartments is guided by ________ _________, which are streches of amino acids usually 15-60 residues long and are localized to N or C terminus or within the protein sequence.
Sorting signals
Multiple, scattered sequences of sorting signals brought together by protein folding are known as a what?
Signal patch
Signal sequences are both necessary and sufficient for protein targeting. They are recognized by _____________ receptors.
Complementary
What removes the sorting signal after a protein reaches its final destination?
Signal peptidase
What type of transport is nuclear transport?
Gated transport, bidirectional and selective
What are the sorting signals called that direct molecules to the nucleus and where are they found?
Nuclear localization signals (NLS), located at many different sites on protein (can be N terminus, C terminus, or within protein sequence)
Nuclear localization signals are recognized by nuclear import receptors (NIRs) or nuclear export receptors (NERs). Which of the following is also an important shuttle for a protein with an NLS?
A. Ran
B. Rab
C. Ras
D. Rho
A. Ran
Ran-GDP and Ran-GTP are utilized in nuclear import and export [note that the GTP form is always within the nucleus]
Describe the directionality of transmembrane transport such as mitochondrial transport
Unidirectional
Most mitochondrial proteins are encoded in ________ DNA
Nuclear
Signal sequences on mitochondrial proteins are located where? - these direct proteins to the correct compartment in mitochondria and specific receptor proteins recognize their configuration. Multi-subunit protein complexes called protein translocators mediate translocation
They are located towards the ends of the proteins to make ampiphilic nuclear structures
What type of mitochondrial membrane translocator transports all nuclear encoded proteins in the outer membrane?
TOM
What type of mitochondrial membrane translocator inserts beta barrel proteins in the outer membrane?
SAM
What type of mitochondrial membrane translocator mediates the insertion of proteins in the inner membrane and matrix?
TIM23
What type of mitochondrial membrane translocator mediates the insertion of transporters in the inner membrane?
TIM22
What type of mitochondrial membrane translocator mediates the insertion of mitochondrially synthesized proteins?
OXA
Transport across the endoplasmic reticulum requires an ER signal sequence guided to the ER membrane by what 2 components?
SRP = signal recognition particle, and an SRP receptor
The SRP utilized in transport across the ER is rod-shaped with a large ___________ pocket lined by methionines.
Hydrophobic
This pocket can accommodate hydrophobic signal sequences of different sizes, shapes, and sequences
What is the role of the ER signal sequence and SRP in directing ribosomes to the ER membrane?
The SRP has the hydrophobic pocket that recognizes proteins supposed to go to the ER. The SRP hugs the ribosome, holds translation (stopping protein synthesis), and brings the entire unit to the ER membrane with another translocator which allows it through the ER membrane and into the lumen
Proteins and other biomolecules are transported via transport vesicles, which bud off from primary compartments and fuse with the next one. They come in different shapes and sizes. The contents of the vesicle are called ________ and transport is __________
Cargo
Directional
There are 3 different vesicular transport systems, what are they?
Biosynthetic - for newly synthesized proteins
Endocytic - proteins coming from outside thru PM into the cell
Retrieval - taking proteins back to previous compartment to maintain homeostasis
The budding off of cargo into a vesicle involves the formation of what at the plasma membrane?
Clathrin coat
The specificity in vesicle targeting is achieved by surface markers on vesicles and complementary receptors on the target membrane. _______ proteins direct vesicles to specific spots on the target membrane, while ________ proteins mediate fusion of the vesicles with the membrane
Rab
SNARE
The sorting of lysosomal proteins requires ________, which is the signal for the protein destined to go to the lysosome
M6P
[recall that this signal must be uncovered by addition of p-GlcNAc]
It can be challenging to isolate a single protein from thousands of others present in a cell. _________ DNA technology can be used to overexpress a protein prior to purification. This allows purification of ________ proteins. A _________________ ______________ typically needs to be done to reduce the complexity of the material prior to purification
Recombinant
Endogenous
Sub-cellular fractionation
During sub-cellular fractionation, a suspension of different cell types is subjected to centrifugation, which separates different cell types based on _______ and ________
Size
Density
What type of centrifugation allows for the separation of organelles?
Ultracentrifugation
What laboratory technique allows for the purification of a single protein from individual organelles?
Column chromatography
There are 3 different matrices for column chromatography, what are they?
Ion-exchange chromatography - separates based on charge
Gel-filtration chromatography - separates based on size
Affinity chromatography - based on specific ligand
What laboratory technique takes a mixture of proteins, applies uniform negative charge, and thus allows you to separate the proteins based on SIZE?
SDS-PAGE
Once you run SDS-PAGE to separate your proteins based on size, what laboratory method would you use to identify a specific protein?
Mass spectrometry
What is the primary use of mass spectrometry?
Identification of unknown proteins
What laboratory technique allows you to analyze specific, known proteins to learn more about them?
Western blotting