CHAPTER ONE

INTRODUCTION

RAC is a subfamily of Rho GTPases (guanosine triphosphatases), which coordinate cellular

response to extracellular signals with the help of highly conserved low molecular weight
proteins that act as molecular switches (Troeger & Williams, 2013).

RAC proteins are categorized into three highly homologous proteins based on their
expression levels—RAC1, RAC2, and RAC3, which play a central role in regulating response to
inflammatory signals mediated by neutrophils, including chemotaxis, actin cytoskeleton
remodeling, and production of superoxide by NADPH oxidase (Dinauer, 2003).

RAC after being activated [guanosine triphosphate (GTP) bound] plays two roles in NADPH
activation which include triggering oxidase complex assembly and functioning as an enzyme
subunit (Babior, 1999; Price, Atkinson, & Knaus, 2002).

Rho GTPases are primarily activated by the VAV family (a group of tyrosine phosphorylation-
regulated signal transduction molecules hierarchically located downstream of protein
tyrosine kinases) of guanine nucleotide exchange factors (GEFs) by catalyzing the exchange of
guanosine diphosphate for GTP.

RAC acts as a molecular switch and remains inactive when bound to Guanosine diphosphate
(GDP). RAC is activated once GEFs remove GDP and permit Guanosine-5'-triphosphate (GTP)
to bind. When bound to GTP, RAC is activated and participates in the regulation of cell
movement. RAC is involved in structural changes to the actin cytoskeleton.

RAC-GTPases facilitate the recruitment of neutrophils to the infected tissues and regulate
degranulation of azurophil and integrin-dependent phagocytosis.

MRC1 (Mannose Receptor C-type 1)is a protein that in humans is encoded by the MRC1 gene.
MRC1 is a type I membrane receptor that mediates the endocytosis of glycoproteins by
macrophages.

MRC1 has been shown to bind high-mannose structures on the surface of potentially
pathogenic viruses, bacteria, and fungi so that they can be neutralized by phagocytic
engulfment.

MRC1 is a significant marker in alternative activation of macrophages in both human and

mice.

MRC1 (Mannose Receptor C-type 1) is a protein that in humans is encoded by the MRC1 gene.
MRC1 is a type I membrane receptor that mediates the endocytosis of glycoproteins by
macrophages. It is also known as CD206.
The Mannose Receptor (Cluster of Differentiation 206, CD206) is a C-type lectin primarily
present on the surface of macrophages, immature dendritic cells and liver sinusoidal
endothelial cells, but is also expressed on the surface of skin cells such as human dermal
fibroblast and keratinocytes.

TYPES OF RAC

RAC (Rho GTPase-activating protein) is a family of proteins that share a common domain
structure and function. There are several types of RAC proteins, including:

1. RAC1 (Ras-related C3 botulinum toxin substrate 1)

2. RAC2 (Ras-related C3 botulinum toxin substrate 2)

3. RAC3 (Ras-related C3 botulinum toxin substrate 3)

4. RAC4 (Ras-related C3 botulinum toxin substrate 4)

These different types of RAC proteins have distinct functions and are involved in various
cellular processes, including:

- Cell migration and adhesion

- Cell proliferation and survival

- Cytoskeletal dynamics

- Signal transduction pathways

- Gene expression regulation

Additionally, there are also different isoforms of RAC proteins, which are generated through
alternative splicing or post-translational modifications. These isoforms may have distinct
functions or subcellular localizations.

It's worth noting that RAC proteins are highly conserved across species, and their functions
are often studied in the context of human disease, including cancer, cardiovascular disease,
and neurological disorders.

TYPES OF MRC1

MRC1 (Mannose Receptor C-type 1) is a protein that has several isoforms and variants,
including:
1. _MRC1a_: The full-length isoform of MRC1, which is widely expressed in various tissues.

2. _MRC1b_: A shorter isoform of MRC1, which is primarily expressed in macrophages and

dendritic cells.

3. _MRC1c_: A variant of MRC1 that is generated through alternative splicing and is expressed
in certain tissues.

4. _sMRC1_: A soluble form of MRC1 that is generated through proteolytic cleavage and is
present in serum and other body fluids.

5. _MRC1-ΔC_: A variant of MRC1 that lacks the cytoplasmic domain and is primarily
expressed in certain cell types.

6. _MRC1-ΔN_: A variant of MRC1 that lacks the extracellular domain and is primarily
expressed in certain cell types.

These different isoforms and variants of MRC1 may have distinct functions, subcellular
localizations, and expression patterns, and may play roles in various cellular processes,
including:

- Endocytosis and phagocytosis

- Antigen presentation and immune response

- Cell adhesion and migration

- Signal transduction pathways

- Tissue development and repair

It's worth noting that the specific functions and characteristics of each MRC1 isoform and
variant may vary depending on the context and cell type.

CHAPTER TWO

LITERATURE REVIEW

2.1. Historical Background and Taxonomy Of RAC

RAC protein has been studied extensively in the context of cancer development and
progression.

Recent experiments on Drosophila suggested that Rac could be involved in mediating the
process of forgetting.

Hyperactivation of Rac increases the memory decay, whereas its inhibition prevents
interference-induced forgetting and slows down a passive memory decay.

RAC is a subfamily of the Rho family of GTPases, small (~21 kDa) signaling G proteins.

The Rho family of GTPases includes Rac, Rho, and Cdc42 small G-protein groups.

Rac comprises Rac1, Rac2, Rac3, and RhoG subgroups.

2.2. Functions Of RAC

RAC proteins are a family of small GTPases that play crucial roles in various cellular
processes, including:

1. Cell migration and adhesion

2. Cell proliferation and survival

3. Cytoskeletal dynamics

4. Membrane trafficking and transport

5. Signal transduction pathways

RAC proteins act as molecular switches, cycling between active (GTP-bound) and inactive
(GDP-bound) states. In their active state, RAC proteins interact with downstream effectors to
regulate various cellular processes.

Some specific functions of RAC proteins include:

1. Regulating actin cytoskeleton dynamics and cell migration

2. Activating protein kinases, such as PAK (p21-activated kinase) and AKT

3. Modulating cell adhesion and junction formation

4. Influencing cell cycle progression and apoptosis

5. Participating in endocytosis and membrane trafficking

6. Interacting with other signaling proteins, such as RHO and CDC42

7. Rac acts as a molecular switch, remaining inactive while bound to GDP and activated once
GEFs remove GDP, permitting GTP to bind.

8. When bound to GTP, Rac is activated and participates in the regulation of cell movement,
through its involvement in structural changes to the actin Cytoskeleton.

9. Rac-GTPases facilitate the recruitment of neutrophils to the infected tissues, and regulate
degranulation of azurophil and integrin-dependent phagocytosis.

10. Rac is required for ROS (reactive oxygen species) production involved in the formation of
NETs (neutrophil extracellular traps), thus facilitating the pathogen and debris clearance by
neutrophils, and the reduction of inflammation.

2.3. Structures Of RAC

RAC proteins are small GTPases that share a conserved structure, consisting of:

1. N-terminal domain ( residues 1-30):

- Contains a guanine nucleotide-binding site

- Involved in protein-protein interactions

2. GTPase domain (residues 31-150):

- Contains the GTP-binding site

- Catalyzes GTP hydrolysis

- Has a characteristic P-loop motif (phosphate-binding loop)

3. Switch I region (residues 151-170):

- Involved in conformational changes upon GTP binding

- Interacts with downstream effectors

4. Switch II region (residues 171-190):

- Also involved in conformational changes

- Interacts with other proteins

5. C-terminal domain (residues 191-206):

- Contains a hypervariable region (HVR)

- Involved in protein-protein interactions and membrane targeting

RAC proteins have a molecular weight of approximately 21 kDa and are composed of 206
amino acids.

The GTPase domain is the most conserved region among RAC proteins, with a high degree of
similarity to other small GTPases. The N-terminal and C-terminal domains are more variable,
allowing for isoform-specific interactions and functions.

The structure of RAC proteins allows them to interact with various proteins and nucleotides,
enabling their role as molecular switches in cellular signaling pathways.

2.4. Historical Background and Taxonomy Of MRC1

History and Development of MRC1

The early history of development of botanical science is nothing but a history of

development of plant taxonomy. The herbalists and agriculturists of ancient times gathered
some knowledge about plants which was passed on from generation to generation.

Theophrastus (372-287 BC), the Greek philosopher-scientist, placed this knowledge of plants
on a scientific footing.

In his “Enquiry into Plants” he dealt with the plants at large and attempted to arrange the
plants in several groups.

He is, therefore, called the “Father of Botany”.

2.5. Classification Of MRC1

1. Linnaeus and the botanists before him tried to classify the plant kingdom using a single or a
few characters chosen arbitrarily.

2. They only thought about the convenience of following a system of classification solely to
identify a particular plant.

3. Such systems are, therefore, called artificial systems of classification.

4. Later on plant-taxonomists conceived the idea that the plants belonged to some natural
groups and they tried to designate and distinguish such groups and tried to classify the plant
kingdom accordingly.

5. Such systems are known as natural systems of classification.

2.6. Structure And Functions Of MRC1

MRC1 (Mannose Receptor C Type 1) is a transmembrane receptor protein that plays a crucial
role in the immune system.
Structure of MRC1

MRC1 is a 175 kDa protein composed of an extracellular domain, a transmembrane domain,

and a cytoplasmic domain. It is primarily expressed on the surface of macrophages, dendritic
cells, and endothelial cells.

Functions of MRC1

1. Role in Immune Response

MRC1 recognizes and binds to mannose-containing glycoproteins on the surface of

microorganisms, such as bacteria, yeast, and viruses. This recognition triggers phagocytosis
and activation of immune responses, including production of cytokines and chemokines.

2. Involvement in Diseases

MRC1 has been implicated in various diseases, including:

- Infections: MRC1 plays a crucial role in host defense against microbial infections.

- Cancer: MRC1 expression is associated with tumor progression and metastasis.

- Autoimmune disorders: MRC1 may contribute to the development of autoimmune diseases,

such as rheumatoid arthritis.

3. Regulation and Signaling

MRC1 signaling involves activation of protein tyrosine kinases, such as Syk and Src, and
recruitment of adaptor proteins, like FcRγ. MRC1 also interacts with other receptors,
including FcγR and CR3, to modulate immune responses.

4. Therapeutic Potential

Targeting MRC1 with monoclonal antibodies or small molecule inhibitors may offer a
promising approach for treating infectious and inflammatory diseases.

CHAPTER THREE

RAC-MRC1: MASTER REPLICATION

RAC and MRC1 are two proteins that interact in cellular signaling, as we discussed earlier.
Replication, in a biological sense, refers to the process of duplicating DNA or other cellular
components. While RAC and MRC1 do play roles in cellular processes, they are not directly
involved in replication.

3.2. Relationship between RAC and MRC1

RAC (Rho-associated coiled-coil containing protein kinase) and MRC1 (Mannose Receptor C-
type 1) have a functional relationship in cellular signaling.

1. *Interaction*: RAC1, a subfamily of RAC proteins, interacts with MRC1, a transmembrane

receptor, to regulate cellular processes.

2. *Phagocytosis*: RAC1 modulates MRC1-mediated phagocytosis of pathogens, apoptotic

cells, and debris.

3. *Cell migration*: RAC1 and MRC1 interact to regulate macrophage migration and adhesion.

4. *Inflammation*: RAC1-MRC1 interaction influences the production of pro-inflammatory

cytokines.

5. *Signaling pathways*: RAC1 activates downstream signaling pathways, including the NF-κB
and MAPK pathways, which regulate various cellular processes, including inflammation and
cell survival.

6. *Regulation of immune response*: The RAC1-MRC1 interaction plays a crucial role in

regulating the immune response, including the activation of macrophages and the production
of cytokines.

RAC-MRC1, also known as Master Replication, refers to the role of the RAC-GTPase protein in
regulating DNA replication. It acts as a master regulator, coordinating and controlling the
replication process in cells.

RAC-MRC1 regulates DNA replication by interacting with various proteins involved in the
replication process. It helps to ensure that DNA is accurately and efficiently duplicated
during cell division. It’s like a conductor guiding the replication orchestra.

RAC-MRC1 interacts with several key proteins involved in DNA replication. One of its
important interactions is with the MCM complex, which is responsible for unwinding the DNA
strands. RAC-MRC1 helps to recruit and activate the MCM complex, ensuring that DNA
replication proceeds smoothly.

RAC(GTPase) has a vital role in DNA replication. It acts as a master regulator, ensuring that the
replication process occurs accurately and efficiently.
By interacting with various proteins involved in replication, RAC(GTPase) helps to coordinate
the unwinding of DNA strands and the recruitment of other essential factors.

RAC(GTPase)-MRC1, the Master of Replication, is a protein that plays a crucial role in

regulating DNA replication. It acts as a master controller, overseeing and coordinating the
replication process in cells.

RAC(GTPase) is a protein that belongs to the Rho family of GTPases. It plays a crucial role in
various cellular processes, including cell migration, cytoskeletal organization, and cell
division. RAC(GTPase) acts as a molecular switch, cycling between an inactive GDP-bound
state and an active GTP-bound state.

When activated, RAC(GTPase) regulates signaling pathways that control cell growth and
proliferation. Its dynamic nature and involvement in multiple cellular functions make
RAC(GTPase) a fascinating protein to study.

RAC(GTPase) is a multitasking protein with various functions in cells. Apart from its role in
DNA replication, it is involved in cell migration, cytoskeletal organization, and cell signaling.
RAC(GTPase) helps cells move and change shape, which is important for processes like wound
healing and immune response.

It also participates in the formation of cellular structures like filopodia and lamellipodia,
which are essential for cell movement. Additionally, RAC(GTPase) is involved in signaling
pathways that regulate cell growth, survival, and differentiation. It’s like a versatile performer,
contributing to different aspects of cellular life!

MRC1, the replication factor. MRC1, also known as Mediator of Replication Checkpoint
protein 1, plays a crucial role in DNA replication. It acts as a checkpoint regulator, ensuring
that DNA replication proceeds accurately and without errors. MRC1 helps to coordinate the
replication machinery and prevents the replication fork from stalling or collapsing.

RAC-MRC1, the dynamic duo of DNA replication! They work together like a well-coordinated
team to ensure accurate and efficient replication.

RAC(GTPase) acts as a master regulator, overseeing the entire process, while MRC1 keeps a
close eye on the replication checkpoint, making sure everything goes smoothly. It’s like a well-
choreographed dance, with each partner playing their part to perfection!

RAC(GTPase) plays a crucial role in coordinating various steps of DNA replication, including
the activation of replication origins and the progression of the replication fork. It ensures that
the replication process is properly regulated and synchronized. On the other hand, MRC1 acts
as a checkpoint protein, monitoring the replication process and preventing any errors or
abnormalities from progressing further.
In summary, the relationship between RAC and MRC1 is one of functional interaction, where
RAC1 regulates MRC1-mediated cellular processes, including phagocytosis, cell migration,
and inflammation, to modulate the immune response and maintain tissue homeostasis.